Whether you’re starting with cultured cells or or tissue samples, or working with plants, bacteria, or mammalian cells, Ambion RNA extraction reagents and kits offer the right product for your research. The Ambion brand includes legacy Invitrogen products such as trusted TRIzol Reagents and PureLink purification kits, alongside premier Ambion products such as Cells-to-CT, MagMAX, and RNAlater kits and reagents.
Which RNA extraction kit from blood sample is right for you?
Order now | Order now | Order now | Order now | Order now | Order now | |
Optimized for liquid samples | Specialized kit deplete non-nucleated erythrocytes | Filter plate format with 96 columns, for HTP needs | HTP optimized for purification from stabilized blood samples | HTP format,capable of recovering RNA of all sizes (including microRNAs) | High RNA yield directly from whole blood, no need to isolate WBCs | |
---|---|---|---|---|---|---|
TRIzol LS | PureLink Total RNA Blood Kit | PureLink Pro 96 total RNA Purification Kit | MagMAX for Stabilized Blood Tubes RNA Isolation Kit | MagMAX mirVana Total RNA Isolation | RiboPure Blood Kit | |
Top Seller | ✓ | |||||
Compatible stabilization reagents | EDTA, Heparin, Citrate, RNAlater | EDTA, Heparin, Citrate | EDTA, Heparin, Citrate | Tempus or PaxGene tubes | EDTA or Citrate | EDTA, Heparin, Citrate, RNAlater |
Stabilization reagent included | ✓ RNAlater | |||||
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Fast, convenient silica columns in a plate format | Scalable, flexible format with magnetic beads | Scalable, flexible format with magnetic beads | Highest purity & convenience; includes both organic extraction & silica column (no precipitation required) |
Prep time | 1 hour | 20 minutes | 12 tubes in 2 hours | 96 samples in <1 hr | 30 minutes | |
High throughput compatible | ✓ | ✓ | ✓ |
For more info on working with blood please review our technical articles:
Supplemental Protocols
- Blood Fractionation Protocol for Collection of White Blood Cells
- Isolation of Small RNAs Using the RiboPure-Blood Kit
Tips from the Bench
TechNotes Articles
- Tips & Tricks for Working with Blood Samples
- Extract MicroRNA from Blood Samples - TechNotes 13(1)
- High Throughput RNA Recovery from Mammalian, Plant, and Viral Samples - TechNotes 13(1)
- Improve RNA Isolation for Array Analysis and More - TechNotes 10(3)
- Improved Gene Expression Profiling With Mouse Blood Samples - TechNotes 13(4)
- Improved Methods for Gene Expression Profiling from Blood Samples - TechNotes 13(3)
- Improved Microarray Sensitivity using Whole Blood RNA Samples - TechNotes 12(3)
- Isolate RNA from White Blood Cells Captured by a Novel Filter System - TechNotes 12(4)
- Low and High Throughput RNA Isolation from Tissues, Cell-free Samples, and Blood - TechNotes 13(2)
- miRNA Expression in White Blood Cells - TechNotes 11(3)
- Obtaining the Highest Quality RNA from Blood Samples - TechNotes 16(1)
- Purify RNA Directly from Whole Blood - TechNotes 11(4)
- Quantitate miRNA from a Few Drops of Mouse Blood - TechNotes 14(4)
- Research Highlight: Superior Microarray Data from Blood Samples - TechNotes 14(4)
- RNA Extraction from Blood: Choosing the Best System for Your Gene Expression Studies - TechNotes 14(2)
- RNA from Mouse Whole Blood for Expression Profiling - TechNotes 13(3)
- Robust High Throughput RNA Isolation from Blood Samples - TechNotes 12(3)
- Superior Gene Expression Profiling with Human Whole Blood Samples - TechNotes 14(3)
- Total RNA from Whole Blood for Expression Profiling - TechNotes 12(2)
Purifying RNA from cultured cells has traditionally been a highly repetitive, time- and labor-intensive procedure, often involving the use of toxic organic chemicals. The products developed by our RNA experts allow you to isolate RNA of maximum yield, purity, and integrity using several technology formats (organic extraction, glass-fiber columns, magnetic beads, and combinations of these).
You can also analyze cultured cell lysates without first isolating the RNA. In significantly less time, you’ll get qPCR results equivalent to - or in some cases better than - those from purified RNA. The Invitrogen products such as Cells-to-CT and Single Cell-to-CT product lines offer tremendous performance and handling advantages.
Which RNA Isolation Kit from Cultured Cells is Right for You?
Order now | Order now | Order now | Order now | Order now | Order now | |
Highly pure, intact RNA | Fast and convenient column based format for RNA (>200 nt) | Fast and convenient column based, filter plateformat for 96 samples (Rna > 200 nt) | Fast and convenient column based method from limited sample input (Rna >200 nt) | Fast and convenient method for small RNA species | Complete system for cells to qRT-PCR without RNA purification | |
TRIzol Reagent | PureLink RNA Mini Kit | PureLink Pro 96 total RNA Purification Kit | PureLink RNA Micro Scale Kit | MirVana miRNA Isolation Kit | Cells-to-CT kits | |
Top seller | ✓ | ✓ | ✓ | ✓ | ||
Prep time | 1 hour | 20 minutes | 20 minutes | 20 minutes | 30 minutes | <10 minutes |
Cell lysis method | High purity organic extraction (requires alcohol precipi-tation) | Nontoxic guanidine-isothio-cyanate lysis | Nontoxic guanidine-isothio-cyanate lysis | Nontoxic guanidine-isothio-cyanate lysis | Organic extraction followed by alcohol precipitation | Cells-to-CT lysis |
Amount of Starting material | Up to 1x107 cells | 5x106 to 1x108 cells | 5x106 to 1x108 cells | Up to 5x105 cells | 102-107 cultured cells | 10-100,000 cells |
Appli-cation | All RNA expression methods | qRT-PCR, NGS | qRT-PCR, NGS | qRT-PCR, NGS | Micro RNA analysis | qRT-PCR |
Preserve and Stabilize Your RNA
RNAlater Solution is widely used for the preservation of RNA in intact animal tissues, organs, and cells, and bacteria. It eliminates the need to immediately process samples for RNA isolation or to freeze them in liquid nitrogen for later processing.
Technical Resources
Which RNA Extraction Kit from FFPE Samples is Right for You?
Order now | Order now | |
Low-throughput isolation of RNA and DNA from the same FFPE section | Scalable isolation of RNA and DNA from the same FFPE section | |
---|---|---|
RecoverAll Total Nucleic Acid Isolation Kit for FFPE | MagMAX FFPE DNA/RNA Ultra Kit | |
Prep Time | 4.5 hours (total time for 16 samples) 2.5 hrs hands-on time | 4 hours (total time for 96 preps) 1 hr hands-on time |
Final product extracted | total RNA, microRNA, gDNA | total RNA, microRNA, gDNA |
Isolation method | column purification | mag bead purification |
Deparaffinization required | ✓ | ✓ |
Automation compatible (post Deparaffinization) | ✓ (Scripts for KF Flex and Duo Prime) | |
Compatible FFPE sample range | up to 4 x 20 micron sections (300 mm2 tissue) | up to 2 x 20 micron sections (300 mm2 tissue) |
Sample volume | ~50µL purified DNA ~50µL purified RNA | ~50uL purified DNA ~50µL purified RNA |
Compatible with Ion AmpliSeq DNA panels (e.g. Cancer Hotspot Panel v2) | ✓ | ✓ |
Compatible with Ion AmpliSeq RNA panels (e.g. RNA cancer and RNA fusion) | ✓ | ✓ |
Why is RNA Isolation From FFPE such a Challenge?
The recovery of quality RNA from FFPE specimens can be quite challenging. The fixation process causes cross-linkage between nucleic acids and proteins, and covalently modifies RNA by the addition of monomethyl groups to the bases. As a result, the molecules are rigid and susceptible to mechanical shearing, and the cross-links may compromise the use of RNA as a substrate for reverse transcription. Therefore, in order to utilize FFPE tissues as a source for gene expression analysis, a reliable method is required for extraction of RNA from the cross-linked matrix.
How can you Increase your Chances of Success With FFPE?
There are a number of factors that can impact the overall quality and yield of RNA isolated from FFPE tissues. Here are recommendations to address several key factors:
- Upstream tissue procurement and tissue specimen preparation—if possible, tissues should be fixed within one hour of surgical resection. Extensive degradation of RNA can occur before completion of the fixation process. The optimal fixation time is 12–24 hours, using neutral-buffered formalin or paraformaldehyde. Fixed tissues should be thoroughly dehydrated prior to the embedding process.
- Block storage—storage of blocks without cut faces, when possible, prevents ongoing damage from exposure to atmospheric oxygen, water, and other environmental factors such as light and infestation (fungus, insects, etc.).
- Tissue type, size, and amount being used for RNA isolation—the recommended tissue thickness is 10–20 µm The number of sections used is determined by the tissue type (which impacts cell density) and surface area (recommended size: 50–300 mm2). Excess starting material can cause filter clogging, resulting in poor yield.
- Excessive amount of paraffin used for embedding tissues—when possible, excess paraffin should be trimmed away prior to starting the purification protocol. For xylene-based purification methods, two xylene treatments at room temperature should be sufficient for complete deparaffinization. If desired, a more rigorous 37–55°C treatment can be performed for up to 30 minutes. After the xylene deparaffinization, it is crucial that the 100% ethanol is completely removed and the pellets are dry after the two 100% ethanol washes. The magnetic bead method employs novel chemistries to deal with the paraffin that limits input to 20 µm sections.
The MagMAX FFPE DNA/RNA Ultra Kits are designed for rapid, efficient, and automatable isolation of total RNA and DNA from FFPE samples using Invitrogen MagMAX magnetic particles. Invitrogen MagMAX FFPE NA kits offer faster workflows and use less toxic reagents, without sacrificing quality in the process. These kits help deliver nucleic acid yield and purity comparable to the best-in-class RecoverAll filter-based system. The MagMAX FFPE NA kits outperform products with similar protocols, and achieve RNA and DNA yields that are comparable with longer, more hazardous, and lower-throughput protocols.
The RecoverAll Total Nucleic Acid Isolation Kit procedure requires about 45 minutes of hands-on time and can easily be completed in less than 1 day when isolating RNA. FFPE samples are deparaffinized using a series of xylene and ethanol washes. Next, they are subjected to a rigorous protease digestion with an incubation time tailored for recovery of either RNA or DNA. The nucleic acids are purified using a rapid glass-filter methodology that includes an on-filter nuclease treatment, and are eluted into either water or the low-salt buffer provided.
Technical Resources
TechNotes Articles
- Nucleic Acid Extraction from FFPE Samples
- Accurate, Sensitive Quantitation of microRNAs from FFPE Tissues - TechNotes 16(1): This study describes the successful isolation and accurate measurement of miRNA expression from formalin- or paraformalin-fixed, paraffin-embedded (FFPE) samples.
- Faster Nucleic Acid Isolation From FFPE Tissues - TechNotes 16(1): Applied Biosystems has improved the protocol for the Invitrogen Ambion RecoverAll Total Nucleic Acid Isolation Kit for FFPE Tissues, resulting in significantly reduced processing time.
- Optimized Gene Expression Analysis from FFPE Samples - TechNotes 14(3): The optimized protocol presented here can be used to reliably quantify mRNA expression levels using RNA isolated from FFPE samples and real-time RT-PCR.
- Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue - TechNotes 12(3): Ambion's new RecoverAll Kit enables researchers to isolate DNA and/or RNA (including microRNA) from FFPE samples. The nucleic acids recovered are ideal for various downstream applications such as microarray analysis, qRT-PCR, and mutation screening.
- MicroRNA Analysis Using RNA Extracted from Matched Formalin-Fixed Paraffin-Embedded (FFPE) and Fresh Frozen Samples
- High correlation of miRNA quantitation data from matched FFPE and snap-frozen tissues using TaqMan MicroRNA Assays
- DNA Genotyping from Human FFPE Samples - Reliable and Reproducible: Application Note
- Optimized Extraction and Quantification of RNA from FFPE Samples for Gene Expression Analyses
- Protocol: FFPE Small RNA Library Preparation for SOLiD Sequencing
- RecoverAll™ Total Nucleic Acid Isolation Kit
Tips from the Bench
Your Data
- Identifying Cancer-Related mRNA Signatures in FFPE Patient Samples: The Yeatman laboratory at the H. Lee Moffitt Cancer Center and Research Institute (Tampa, FL) investigates the gene expression signatures that define colon, breast, and other cancers.
- LCM Staining Effects on Real-Time RT-PCR: Research is described demonstrating that the Ambion LCM Staining Kit can be successfully used on formalin-fixed and paraffin-embedded (FFPE) tissues without compromising either the yield or the quality of RNA extracted.
Application Note
Product Bulletin
Which Plant RNA Isolation Kit is Right for You?
Order now | Order now | Order now | Order now | |
Great for difficult samples (conifer tissue & seeds) | Quick & easy to use | Efficient recovery of microRNA & small RNA | Rapid & fully automated | |
---|---|---|---|---|
Plant RNA Reagent | PureLink RNA Mini Kit | mirVana miRNA Isolation Kit | MagMAX-96 Total RNA Isolation Kit | |
Top seller | ✓ | |||
RNA types isolated | large RNA molecules only | large RNA molecules only | Small & large RNA molecules | large RNA molecules only |
Prep time | 60 minutes | <20 minutes | 30 minutes | <45 minutes |
Amount of starting material | Up to 1 g | <250 mg | 0.5 to 250 mg | Up to 10 mg |
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Highest purity & convenience; includes organic extraction & silica column (no precipitation required) | Scalable, flexible format with magnetic beads |
High throughput compatible | ✓ | |||
Compatible products | Ambion Plant RNA Isolation Aid Removes common plant contaminants | Ambion Plant RNA Isolation Aid Removes common plant contaminants | Ambion Plant RNA Isolation Aid Removes common plant contaminants |
Remove Polysaccharides
One of the major issues with plant RNA isolation is the presence of problematic biomolecules, such as polyphenolic compounds and polysaccharides. The Ambion Plant RNA Isolation Aid minimizes many of these contaminants with the addition of polyvinylpyrrolidone, a high molecular weight polymer that binds to polyphenolics and polysaccharides and a pre-extraction spin step.
Preserve and Stabilize Your RNA
RNAlater Solution is widely for the preservation of RNA in intact animal tissues, organs, and cells, and bacteria. It minimizes the need to immediately process samples for RNA isolation or to freeze them in liquid nitrogen for later processing.
Technical Resources
Which tissue RNA extraction kit is right for you?
Order now | Order now | Order now | Order now | Order now | Order now | Order now | |
Gold standard for highly pure, intact RNA | Simple, reliable, rapid method | Fast and convenient column based, filter plateformat for 96 samples (Rna > 200 nt) | Fast and convenient column based method from limited sample input (Rna >200 nt) | Fast and convenient method for small RNA species | Highest purity TRIzol + column purification, no precipitation required | HTP format, capable of recovering RNA of all sizes (including microRNAs) | |
---|---|---|---|---|---|---|---|
TRIzol Reagent | PureLink RNA Mini Kit | PureLink Pro 96 total RNA Purification Kit | PureLink RNA Micro Scale Kit | mirVana miRNA Isolation Kit | TRIzol Plus RNA Purification System | MagMAX mirVana Total RNA Isolation | |
Top Seller | ✓ | ||||||
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Fast Convenient silica column in a filter plate format | Fast Convenient micro silica columns | Organic extraction combined with convenient silica columns | Highest purity and convenience; includes both organic extraction and silica column (no precipitation required) | Scalable, flexible format with magnetic beads |
Prep time | 1 hour | 20 minutes | 20 minutes | 20 minutes | 30 minutes | 1 hour | <1 hour |
Amount of starting material | Up to 100 mg | 10 mg to 100 mg | 10 cells to 10 mg | up to 10 mg | up to 250 mg tissue | Up to 100 mg | Up to 50 mg |
High throughput compatible | ✓ | ✓ | |||||
Prep size | 100 preps | 50 preps | 96 preps | 50 preps | 96 preps | 50 preps | 96 preps |
RNA Yields From Animal Tissues
RNA content can vary widely between tissues, cell types, physiological states, etc. This chart provides general guidelines for estimating RNA yields from a variety of cells and tissues.
Table 1. Typical total RNA yields from 1 mg of various tissues.
Preserve and Stabilize Your RNA
RNAlater Solution is widely used for the preservation of RNA in intact animal tissues, organs, and cells, and bacteria. It eliminates the need to immediately process samples for RNA isolation or to freeze them in liquid nitrogen for later processing.
Technical Resources
Which Yeast RNA Extraction Kit is Right for You?
Order now | Order now | Order now | Order now | |
Rapid & fully automated | Quick & easy to use | Designed for rigid yeast cell walls while maintaining expression profiles | Fast and convenient column based, filter plateformat for 96 samples (Rna > 200 nt) | |
---|---|---|---|---|
Platinum Taq MagMAX for Microarrays Total RNA Isolation Kit | PureLink RNA Mini Kit | RiboPure-Yeast Kit | PureLink Pro 96 total RNA Purification Kit | |
Top seller | ✓ | |||
Includes specialized beads for cell lysis | No | No | Zirconia beads | No |
Isolation method | Scalable, flexible format for highest purity; includes organic extraction and magnetic beads | Fast, convenient silica column | Highest purity and convenience; includes both organic extraction and silica column | Fast, convenient silica column in 96 well filter plate format |
Prep time | <1 hour | 20 minutes | 90 minutes (includes DNase treatment) | 20 minutes |
Amount of starting material | Up to 5 x 106 cells | Up to 5 x 108 cells | Up to 3 x 108 cells | 1.8 mL of fresh, log-phase yeast cells (OD660=0.6–0.8) |
High throughput compatible | ✓ | |||
Kit size | 96 preps | 50 preps | 50 preps | 96 preps |
Technical Resources
Unlike eukaryotic cells, yeast and many bacterial strains are protected by cell walls that can be difficult to disrupt. As a first step, effective cell lysis (typically via mechanical disruption or enzymatic pretreatment) is required prior to any RNA isolation procedure.
A range of products developed by the Ambion RNA experts will help you isolate high yields of pure, intact RNA from bacteria and yeast.
Which Bacterial RNA Extraction Kit is Right for You?
Order now | Order now | Order now | Order now | Order now | |
No lysis procedures | Fast & easy, 20 min | Fast and convenient silica column based , filter plate format | HTP, 45 min | Ideal for hard-to-lyse gram + bacteria | |
---|---|---|---|---|---|
TRIzol Max Bacterial RNA Isolation Kit w/TRIzol & Max Bacterial Enhancement Reagent | PureLink RNA Mini Kit | PureLink Pro 96 total RNA Purification Kit | MagMAX Total Nucleic Acid Isolation Kit | RiboPure-Bacteria Kit | |
Top Seller | ✓ | ||||
Includes specialized beads for cell lysis | No | No | No | Zirconia beads | Zirconia beads |
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Fast, convenient silica column in 96 well filter plate format | Scalable, flexible format with magnetic beads | Highest purity & convenience; includes organic extraction & silica column (no precipitation required) |
Prep time | 1 hour | 20 minutes | 20 minutes | 45 minutes | 1 hour |
Amount of starting material | Up to 1 x 108 cells | Up to 1 x 109 cells | Up to 1 x 109 cells | Up to 0.3 g solid samples or 175 µl liquid samples | Up to 109 cells |
High throughput compatible | ✓ | ✓ | |||
Kit size | 100 preps | 50 preps | 96 preps | 100 preps | 50 preps |
Technical Resources
Viral analysis of biological (animal, insect, plant, fungal, bacterial) and environmental (water, air, food) samples is increasingly conducted using molecular testing strategies that require viral RNA or DNA from the samples being tested. Purification of viral RNA poses unique challenges for getting good recovery and detection sensitivity. Ambion RNA experts have developed RNA purification products that are optimized to provide maximum viral RNA yield, purity, and integrity from a broad range of sample types in several format options.
"Highly concentrated viral RNA from the PureLink Viral RNA/DNA Mini Kit provides a tool for more sensitive and delicate downstream analysis on viral genomes”
Product Specifications
Most Suitable Downstream Application | ||||||
---|---|---|---|---|---|---|
Product | Format | Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning |
Total RNA | ||||||
PureLink Viral RNA⁄DNA Mini Kit | Chaotropic salt-based lysis, silica filter | ✓ | ✓ | |||
RNAqueous Kit | Chaotropic salt-based lysis, silica filter | ✓ | ✓ | ✓ | ||
MagMAX AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | ✓ | ✓ | |||
MagMAX-96 AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ✓ | |||
DNA/RNA | ||||||
PureLink Pro 96 Viral RNA⁄DNA Purification Kit | Chaotropic salt-based lysis, silica filter, 96-well processing, automation compatible | ✓ | ||||
MagMAX Total Nucleic Acid Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ✓ | |||
MagMAX-96 Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ✓ | |||
Dynabeads SILANE Viral NA | Chaotropic salt-based lysis, magnetic beads, automation compatible | |||||
iPrep PureLink Virus Kit | Chaotropic salt-based lysis, magnetic particles, automation compatible | ✓ | ||||
MagMAX Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | ✓ | ||||
MagMAX-96 Viral RNA Isolation Kit (5 x 96 rxns) | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ||||
MicroRNA | ||||||
mirVana miRNA Isolation Kit | Chaotropic salt-based lysis, silica filter | ✓ | ||||
mirVana miRNA Isolation Kit (without phenol - must be purchased separately) | Chaotropic salt-based lysis, silica filter | ✓ | ||||
TRIzol LS Reagent | Lysis in TRIzol reagent, one-step reagent | ✓ | ✓ | ✓ | ✓ | |
mRNA | ||||||
Dynabeads mRNA DIRECT Kit | Chaotropic salt-based lysis, magnetic beads | ✓ | ✓ | |||
Dynabeads mRNA DIRECT Micro Kit | Chaotropic salt-based lysis, magnetic beads | ✓ | ✓ |
Which RNA extraction kit from blood sample is right for you?
Order now | Order now | Order now | Order now | Order now | Order now | |
Optimized for liquid samples | Specialized kit deplete non-nucleated erythrocytes | Filter plate format with 96 columns, for HTP needs | HTP optimized for purification from stabilized blood samples | HTP format,capable of recovering RNA of all sizes (including microRNAs) | High RNA yield directly from whole blood, no need to isolate WBCs | |
---|---|---|---|---|---|---|
TRIzol LS | PureLink Total RNA Blood Kit | PureLink Pro 96 total RNA Purification Kit | MagMAX for Stabilized Blood Tubes RNA Isolation Kit | MagMAX mirVana Total RNA Isolation | RiboPure Blood Kit | |
Top Seller | ✓ | |||||
Compatible stabilization reagents | EDTA, Heparin, Citrate, RNAlater | EDTA, Heparin, Citrate | EDTA, Heparin, Citrate | Tempus or PaxGene tubes | EDTA or Citrate | EDTA, Heparin, Citrate, RNAlater |
Stabilization reagent included | ✓ RNAlater | |||||
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Fast, convenient silica columns in a plate format | Scalable, flexible format with magnetic beads | Scalable, flexible format with magnetic beads | Highest purity & convenience; includes both organic extraction & silica column (no precipitation required) |
Prep time | 1 hour | 20 minutes | 12 tubes in 2 hours | 96 samples in <1 hr | 30 minutes | |
High throughput compatible | ✓ | ✓ | ✓ |
For more info on working with blood please review our technical articles:
Supplemental Protocols
- Blood Fractionation Protocol for Collection of White Blood Cells
- Isolation of Small RNAs Using the RiboPure-Blood Kit
Tips from the Bench
TechNotes Articles
- Tips & Tricks for Working with Blood Samples
- Extract MicroRNA from Blood Samples - TechNotes 13(1)
- High Throughput RNA Recovery from Mammalian, Plant, and Viral Samples - TechNotes 13(1)
- Improve RNA Isolation for Array Analysis and More - TechNotes 10(3)
- Improved Gene Expression Profiling With Mouse Blood Samples - TechNotes 13(4)
- Improved Methods for Gene Expression Profiling from Blood Samples - TechNotes 13(3)
- Improved Microarray Sensitivity using Whole Blood RNA Samples - TechNotes 12(3)
- Isolate RNA from White Blood Cells Captured by a Novel Filter System - TechNotes 12(4)
- Low and High Throughput RNA Isolation from Tissues, Cell-free Samples, and Blood - TechNotes 13(2)
- miRNA Expression in White Blood Cells - TechNotes 11(3)
- Obtaining the Highest Quality RNA from Blood Samples - TechNotes 16(1)
- Purify RNA Directly from Whole Blood - TechNotes 11(4)
- Quantitate miRNA from a Few Drops of Mouse Blood - TechNotes 14(4)
- Research Highlight: Superior Microarray Data from Blood Samples - TechNotes 14(4)
- RNA Extraction from Blood: Choosing the Best System for Your Gene Expression Studies - TechNotes 14(2)
- RNA from Mouse Whole Blood for Expression Profiling - TechNotes 13(3)
- Robust High Throughput RNA Isolation from Blood Samples - TechNotes 12(3)
- Superior Gene Expression Profiling with Human Whole Blood Samples - TechNotes 14(3)
- Total RNA from Whole Blood for Expression Profiling - TechNotes 12(2)
Purifying RNA from cultured cells has traditionally been a highly repetitive, time- and labor-intensive procedure, often involving the use of toxic organic chemicals. The products developed by our RNA experts allow you to isolate RNA of maximum yield, purity, and integrity using several technology formats (organic extraction, glass-fiber columns, magnetic beads, and combinations of these).
You can also analyze cultured cell lysates without first isolating the RNA. In significantly less time, you’ll get qPCR results equivalent to - or in some cases better than - those from purified RNA. The Invitrogen products such as Cells-to-CT and Single Cell-to-CT product lines offer tremendous performance and handling advantages.
Which RNA Isolation Kit from Cultured Cells is Right for You?
Order now | Order now | Order now | Order now | Order now | Order now | |
Highly pure, intact RNA | Fast and convenient column based format for RNA (>200 nt) | Fast and convenient column based, filter plateformat for 96 samples (Rna > 200 nt) | Fast and convenient column based method from limited sample input (Rna >200 nt) | Fast and convenient method for small RNA species | Complete system for cells to qRT-PCR without RNA purification | |
TRIzol Reagent | PureLink RNA Mini Kit | PureLink Pro 96 total RNA Purification Kit | PureLink RNA Micro Scale Kit | MirVana miRNA Isolation Kit | Cells-to-CT kits | |
Top seller | ✓ | ✓ | ✓ | ✓ | ||
Prep time | 1 hour | 20 minutes | 20 minutes | 20 minutes | 30 minutes | <10 minutes |
Cell lysis method | High purity organic extraction (requires alcohol precipi-tation) | Nontoxic guanidine-isothio-cyanate lysis | Nontoxic guanidine-isothio-cyanate lysis | Nontoxic guanidine-isothio-cyanate lysis | Organic extraction followed by alcohol precipitation | Cells-to-CT lysis |
Amount of Starting material | Up to 1x107 cells | 5x106 to 1x108 cells | 5x106 to 1x108 cells | Up to 5x105 cells | 102-107 cultured cells | 10-100,000 cells |
Appli-cation | All RNA expression methods | qRT-PCR, NGS | qRT-PCR, NGS | qRT-PCR, NGS | Micro RNA analysis | qRT-PCR |
Preserve and Stabilize Your RNA
RNAlater Solution is widely used for the preservation of RNA in intact animal tissues, organs, and cells, and bacteria. It eliminates the need to immediately process samples for RNA isolation or to freeze them in liquid nitrogen for later processing.
Technical Resources
Which RNA Extraction Kit from FFPE Samples is Right for You?
Order now | Order now | |
Low-throughput isolation of RNA and DNA from the same FFPE section | Scalable isolation of RNA and DNA from the same FFPE section | |
---|---|---|
RecoverAll Total Nucleic Acid Isolation Kit for FFPE | MagMAX FFPE DNA/RNA Ultra Kit | |
Prep Time | 4.5 hours (total time for 16 samples) 2.5 hrs hands-on time | 4 hours (total time for 96 preps) 1 hr hands-on time |
Final product extracted | total RNA, microRNA, gDNA | total RNA, microRNA, gDNA |
Isolation method | column purification | mag bead purification |
Deparaffinization required | ✓ | ✓ |
Automation compatible (post Deparaffinization) | ✓ (Scripts for KF Flex and Duo Prime) | |
Compatible FFPE sample range | up to 4 x 20 micron sections (300 mm2 tissue) | up to 2 x 20 micron sections (300 mm2 tissue) |
Sample volume | ~50µL purified DNA ~50µL purified RNA | ~50uL purified DNA ~50µL purified RNA |
Compatible with Ion AmpliSeq DNA panels (e.g. Cancer Hotspot Panel v2) | ✓ | ✓ |
Compatible with Ion AmpliSeq RNA panels (e.g. RNA cancer and RNA fusion) | ✓ | ✓ |
Why is RNA Isolation From FFPE such a Challenge?
The recovery of quality RNA from FFPE specimens can be quite challenging. The fixation process causes cross-linkage between nucleic acids and proteins, and covalently modifies RNA by the addition of monomethyl groups to the bases. As a result, the molecules are rigid and susceptible to mechanical shearing, and the cross-links may compromise the use of RNA as a substrate for reverse transcription. Therefore, in order to utilize FFPE tissues as a source for gene expression analysis, a reliable method is required for extraction of RNA from the cross-linked matrix.
How can you Increase your Chances of Success With FFPE?
There are a number of factors that can impact the overall quality and yield of RNA isolated from FFPE tissues. Here are recommendations to address several key factors:
- Upstream tissue procurement and tissue specimen preparation—if possible, tissues should be fixed within one hour of surgical resection. Extensive degradation of RNA can occur before completion of the fixation process. The optimal fixation time is 12–24 hours, using neutral-buffered formalin or paraformaldehyde. Fixed tissues should be thoroughly dehydrated prior to the embedding process.
- Block storage—storage of blocks without cut faces, when possible, prevents ongoing damage from exposure to atmospheric oxygen, water, and other environmental factors such as light and infestation (fungus, insects, etc.).
- Tissue type, size, and amount being used for RNA isolation—the recommended tissue thickness is 10–20 µm The number of sections used is determined by the tissue type (which impacts cell density) and surface area (recommended size: 50–300 mm2). Excess starting material can cause filter clogging, resulting in poor yield.
- Excessive amount of paraffin used for embedding tissues—when possible, excess paraffin should be trimmed away prior to starting the purification protocol. For xylene-based purification methods, two xylene treatments at room temperature should be sufficient for complete deparaffinization. If desired, a more rigorous 37–55°C treatment can be performed for up to 30 minutes. After the xylene deparaffinization, it is crucial that the 100% ethanol is completely removed and the pellets are dry after the two 100% ethanol washes. The magnetic bead method employs novel chemistries to deal with the paraffin that limits input to 20 µm sections.
The MagMAX FFPE DNA/RNA Ultra Kits are designed for rapid, efficient, and automatable isolation of total RNA and DNA from FFPE samples using Invitrogen MagMAX magnetic particles. Invitrogen MagMAX FFPE NA kits offer faster workflows and use less toxic reagents, without sacrificing quality in the process. These kits help deliver nucleic acid yield and purity comparable to the best-in-class RecoverAll filter-based system. The MagMAX FFPE NA kits outperform products with similar protocols, and achieve RNA and DNA yields that are comparable with longer, more hazardous, and lower-throughput protocols.
The RecoverAll Total Nucleic Acid Isolation Kit procedure requires about 45 minutes of hands-on time and can easily be completed in less than 1 day when isolating RNA. FFPE samples are deparaffinized using a series of xylene and ethanol washes. Next, they are subjected to a rigorous protease digestion with an incubation time tailored for recovery of either RNA or DNA. The nucleic acids are purified using a rapid glass-filter methodology that includes an on-filter nuclease treatment, and are eluted into either water or the low-salt buffer provided.
Technical Resources
TechNotes Articles
- Nucleic Acid Extraction from FFPE Samples
- Accurate, Sensitive Quantitation of microRNAs from FFPE Tissues - TechNotes 16(1): This study describes the successful isolation and accurate measurement of miRNA expression from formalin- or paraformalin-fixed, paraffin-embedded (FFPE) samples.
- Faster Nucleic Acid Isolation From FFPE Tissues - TechNotes 16(1): Applied Biosystems has improved the protocol for the Invitrogen Ambion RecoverAll Total Nucleic Acid Isolation Kit for FFPE Tissues, resulting in significantly reduced processing time.
- Optimized Gene Expression Analysis from FFPE Samples - TechNotes 14(3): The optimized protocol presented here can be used to reliably quantify mRNA expression levels using RNA isolated from FFPE samples and real-time RT-PCR.
- Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue - TechNotes 12(3): Ambion's new RecoverAll Kit enables researchers to isolate DNA and/or RNA (including microRNA) from FFPE samples. The nucleic acids recovered are ideal for various downstream applications such as microarray analysis, qRT-PCR, and mutation screening.
- MicroRNA Analysis Using RNA Extracted from Matched Formalin-Fixed Paraffin-Embedded (FFPE) and Fresh Frozen Samples
- High correlation of miRNA quantitation data from matched FFPE and snap-frozen tissues using TaqMan MicroRNA Assays
- DNA Genotyping from Human FFPE Samples - Reliable and Reproducible: Application Note
- Optimized Extraction and Quantification of RNA from FFPE Samples for Gene Expression Analyses
- Protocol: FFPE Small RNA Library Preparation for SOLiD Sequencing
- RecoverAll™ Total Nucleic Acid Isolation Kit
Tips from the Bench
Your Data
- Identifying Cancer-Related mRNA Signatures in FFPE Patient Samples: The Yeatman laboratory at the H. Lee Moffitt Cancer Center and Research Institute (Tampa, FL) investigates the gene expression signatures that define colon, breast, and other cancers.
- LCM Staining Effects on Real-Time RT-PCR: Research is described demonstrating that the Ambion LCM Staining Kit can be successfully used on formalin-fixed and paraffin-embedded (FFPE) tissues without compromising either the yield or the quality of RNA extracted.
Application Note
Product Bulletin
Which Plant RNA Isolation Kit is Right for You?
Order now | Order now | Order now | Order now | |
Great for difficult samples (conifer tissue & seeds) | Quick & easy to use | Efficient recovery of microRNA & small RNA | Rapid & fully automated | |
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Plant RNA Reagent | PureLink RNA Mini Kit | mirVana miRNA Isolation Kit | MagMAX-96 Total RNA Isolation Kit | |
Top seller | ✓ | |||
RNA types isolated | large RNA molecules only | large RNA molecules only | Small & large RNA molecules | large RNA molecules only |
Prep time | 60 minutes | <20 minutes | 30 minutes | <45 minutes |
Amount of starting material | Up to 1 g | <250 mg | 0.5 to 250 mg | Up to 10 mg |
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Highest purity & convenience; includes organic extraction & silica column (no precipitation required) | Scalable, flexible format with magnetic beads |
High throughput compatible | ✓ | |||
Compatible products | Ambion Plant RNA Isolation Aid Removes common plant contaminants | Ambion Plant RNA Isolation Aid Removes common plant contaminants | Ambion Plant RNA Isolation Aid Removes common plant contaminants |
Remove Polysaccharides
One of the major issues with plant RNA isolation is the presence of problematic biomolecules, such as polyphenolic compounds and polysaccharides. The Ambion Plant RNA Isolation Aid minimizes many of these contaminants with the addition of polyvinylpyrrolidone, a high molecular weight polymer that binds to polyphenolics and polysaccharides and a pre-extraction spin step.
Preserve and Stabilize Your RNA
RNAlater Solution is widely for the preservation of RNA in intact animal tissues, organs, and cells, and bacteria. It minimizes the need to immediately process samples for RNA isolation or to freeze them in liquid nitrogen for later processing.
Technical Resources
Which tissue RNA extraction kit is right for you?
Order now | Order now | Order now | Order now | Order now | Order now | Order now | |
Gold standard for highly pure, intact RNA | Simple, reliable, rapid method | Fast and convenient column based, filter plateformat for 96 samples (Rna > 200 nt) | Fast and convenient column based method from limited sample input (Rna >200 nt) | Fast and convenient method for small RNA species | Highest purity TRIzol + column purification, no precipitation required | HTP format, capable of recovering RNA of all sizes (including microRNAs) | |
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TRIzol Reagent | PureLink RNA Mini Kit | PureLink Pro 96 total RNA Purification Kit | PureLink RNA Micro Scale Kit | mirVana miRNA Isolation Kit | TRIzol Plus RNA Purification System | MagMAX mirVana Total RNA Isolation | |
Top Seller | ✓ | ||||||
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Fast Convenient silica column in a filter plate format | Fast Convenient micro silica columns | Organic extraction combined with convenient silica columns | Highest purity and convenience; includes both organic extraction and silica column (no precipitation required) | Scalable, flexible format with magnetic beads |
Prep time | 1 hour | 20 minutes | 20 minutes | 20 minutes | 30 minutes | 1 hour | <1 hour |
Amount of starting material | Up to 100 mg | 10 mg to 100 mg | 10 cells to 10 mg | up to 10 mg | up to 250 mg tissue | Up to 100 mg | Up to 50 mg |
High throughput compatible | ✓ | ✓ | |||||
Prep size | 100 preps | 50 preps | 96 preps | 50 preps | 96 preps | 50 preps | 96 preps |
RNA Yields From Animal Tissues
RNA content can vary widely between tissues, cell types, physiological states, etc. This chart provides general guidelines for estimating RNA yields from a variety of cells and tissues.
Table 1. Typical total RNA yields from 1 mg of various tissues.
Preserve and Stabilize Your RNA
RNAlater Solution is widely used for the preservation of RNA in intact animal tissues, organs, and cells, and bacteria. It eliminates the need to immediately process samples for RNA isolation or to freeze them in liquid nitrogen for later processing.
Technical Resources
Which Yeast RNA Extraction Kit is Right for You?
Order now | Order now | Order now | Order now | |
Rapid & fully automated | Quick & easy to use | Designed for rigid yeast cell walls while maintaining expression profiles | Fast and convenient column based, filter plateformat for 96 samples (Rna > 200 nt) | |
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Platinum Taq MagMAX for Microarrays Total RNA Isolation Kit | PureLink RNA Mini Kit | RiboPure-Yeast Kit | PureLink Pro 96 total RNA Purification Kit | |
Top seller | ✓ | |||
Includes specialized beads for cell lysis | No | No | Zirconia beads | No |
Isolation method | Scalable, flexible format for highest purity; includes organic extraction and magnetic beads | Fast, convenient silica column | Highest purity and convenience; includes both organic extraction and silica column | Fast, convenient silica column in 96 well filter plate format |
Prep time | <1 hour | 20 minutes | 90 minutes (includes DNase treatment) | 20 minutes |
Amount of starting material | Up to 5 x 106 cells | Up to 5 x 108 cells | Up to 3 x 108 cells | 1.8 mL of fresh, log-phase yeast cells (OD660=0.6–0.8) |
High throughput compatible | ✓ | |||
Kit size | 96 preps | 50 preps | 50 preps | 96 preps |
Technical Resources
Unlike eukaryotic cells, yeast and many bacterial strains are protected by cell walls that can be difficult to disrupt. As a first step, effective cell lysis (typically via mechanical disruption or enzymatic pretreatment) is required prior to any RNA isolation procedure.
A range of products developed by the Ambion RNA experts will help you isolate high yields of pure, intact RNA from bacteria and yeast.
Which Bacterial RNA Extraction Kit is Right for You?
Order now | Order now | Order now | Order now | Order now | |
No lysis procedures | Fast & easy, 20 min | Fast and convenient silica column based , filter plate format | HTP, 45 min | Ideal for hard-to-lyse gram + bacteria | |
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TRIzol Max Bacterial RNA Isolation Kit w/TRIzol & Max Bacterial Enhancement Reagent | PureLink RNA Mini Kit | PureLink Pro 96 total RNA Purification Kit | MagMAX Total Nucleic Acid Isolation Kit | RiboPure-Bacteria Kit | |
Top Seller | ✓ | ||||
Includes specialized beads for cell lysis | No | No | No | Zirconia beads | Zirconia beads |
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Fast, convenient silica column in 96 well filter plate format | Scalable, flexible format with magnetic beads | Highest purity & convenience; includes organic extraction & silica column (no precipitation required) |
Prep time | 1 hour | 20 minutes | 20 minutes | 45 minutes | 1 hour |
Amount of starting material | Up to 1 x 108 cells | Up to 1 x 109 cells | Up to 1 x 109 cells | Up to 0.3 g solid samples or 175 µl liquid samples | Up to 109 cells |
High throughput compatible | ✓ | ✓ | |||
Kit size | 100 preps | 50 preps | 96 preps | 100 preps | 50 preps |
Technical Resources
Viral analysis of biological (animal, insect, plant, fungal, bacterial) and environmental (water, air, food) samples is increasingly conducted using molecular testing strategies that require viral RNA or DNA from the samples being tested. Purification of viral RNA poses unique challenges for getting good recovery and detection sensitivity. Ambion RNA experts have developed RNA purification products that are optimized to provide maximum viral RNA yield, purity, and integrity from a broad range of sample types in several format options.
"Highly concentrated viral RNA from the PureLink Viral RNA/DNA Mini Kit provides a tool for more sensitive and delicate downstream analysis on viral genomes”
Product Specifications
Most Suitable Downstream Application | ||||||
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Product | Format | Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning |
Total RNA | ||||||
PureLink Viral RNA⁄DNA Mini Kit | Chaotropic salt-based lysis, silica filter | ✓ | ✓ | |||
RNAqueous Kit | Chaotropic salt-based lysis, silica filter | ✓ | ✓ | ✓ | ||
MagMAX AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | ✓ | ✓ | |||
MagMAX-96 AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ✓ | |||
DNA/RNA | ||||||
PureLink Pro 96 Viral RNA⁄DNA Purification Kit | Chaotropic salt-based lysis, silica filter, 96-well processing, automation compatible | ✓ | ||||
MagMAX Total Nucleic Acid Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ✓ | |||
MagMAX-96 Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ✓ | |||
Dynabeads SILANE Viral NA | Chaotropic salt-based lysis, magnetic beads, automation compatible | |||||
iPrep PureLink Virus Kit | Chaotropic salt-based lysis, magnetic particles, automation compatible | ✓ | ||||
MagMAX Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | ✓ | ||||
MagMAX-96 Viral RNA Isolation Kit (5 x 96 rxns) | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | ✓ | ||||
MicroRNA | ||||||
mirVana miRNA Isolation Kit | Chaotropic salt-based lysis, silica filter | ✓ | ||||
mirVana miRNA Isolation Kit (without phenol - must be purchased separately) | Chaotropic salt-based lysis, silica filter | ✓ | ||||
TRIzol LS Reagent | Lysis in TRIzol reagent, one-step reagent | ✓ | ✓ | ✓ | ✓ | |
mRNA | ||||||
Dynabeads mRNA DIRECT Kit | Chaotropic salt-based lysis, magnetic beads | ✓ | ✓ | |||
Dynabeads mRNA DIRECT Micro Kit | Chaotropic salt-based lysis, magnetic beads | ✓ | ✓ |
Resources
Support
For Research Use Only. Not for use in diagnostic procedures.