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Custom TaqMan™ Small RNA Assay Applied Biosystems™

Custom TaqMan Small RNA Assays use TaqMan Real-Time PCR Assays for the analysis of any small RNA molecule. The novel adaptations in TaqMan Assay design developed for the study of miRNAs are ideal for analysis of any small nucleic acid less than 200 bases long, including newly discovered miRNAs, Piwi-interacting RNA (piRNA), small nuclear RNA (snRNA), and small nucleolar RNA (snoRNA).

Flexible and convenient ordering—easy process with our secure Custom TaqMan Small RNA Assay design tool
Versatile application—use for the analysis of any small RNA from any species
Reliable outcome—pre-formulated assays are optimized to run under universal conditions
Powerful miRNA quantitation—the same sensitivity, specificity, and reproducibility as predesigned TaqMan MicroRNA Assays

Sophisticated design pipeline
The miRPipe Small RNA Assay design pipeline is founded on our extensive background and bioinformatics expertise designing TaqMan Gene Expression Assays. To detect very small targets, it includes design features based on empirical data gathered over years of experiments. The resulting automated pipeline includes built-in flexibility features enabling design of assays for the broadest assortment of small RNA sequences. The benefits of TaqMan Assay technology are ideal for analysis of virtually any small RNA, enabling results you can trust from the very first experiment.

Ordering information
First, bioinformatically qualify your sequence target for specificity and sequence quality, then use the Custom TaqMan Small RNA Assay Design Tool to input your sequence and submit it for assay design. This easy-to-use tool is a secure and confidential. If we are unable to design or manufacture an assay for your sequence target of interest, you will not be charged.

Purification , Cartridge , 1 µmol Invitrogen™

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3' and 5' modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE).

TaqMan™ Gene Expression Cells-to-CT™ Kit Invitrogen™

The TaqMan® Gene Expression Cells-to-CT™ Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification. This kit saves time and offers a simple workflow that is suitable for a few samples or can be easily incorporated into automated, high-throughput applications.

The TaqMan® Gene Expression Cells-to-CT Kit is:

• Complete—optimized workflow includes cell lysis reagents with gDNA removal, RT enzyme mix, buffer, and new TaqMan® Gene Expression Master Mix
• Fast—7-minute sample prep, including DNase treatment, at room temperature
• Easy—lyse samples in a tube or directly in culture plates
• Robust—perform gene expression analysis on 10–100,000 cells per sample; results equivalent to those from purified RNA
• Efficient—contains sufficient reagents to generate 500 real-time PCR results from 100 starting samples

Featuring a unique method for lysing cultured cells while removing genomic DNA and preserving RNA integrity, the TaqMan® Gene Expression Cells-to-CT™ Kit contains reverse transcription (RT) reagents for cDNA synthesis and TaqMan® Gene Expression Master Mix for real-time PCR analysis. TaqMan® primer⁄probe sets are sold separately.

Simple 7-minute sample preparation: part of a complete workflow
Whether you are using plates or tubes, the TaqMan® Gene Expression Cells-to-CT™ Kit, which uses the simple 7-minute sample preparation procedure outlined (see figure), is designed for 10–100,000 cultured cells⁄sample. Cells are washed in PBS and lysed in solution for 5 minutes at room temperature; DNase treatment can be performed simultaneously. Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution. The lysates are now ready for reverse transcription or storage at –20°C. Because samples can be processed directly in culture wells (96 or 384 wells), sample handling and the potential for sample loss or transfer error are minimized, facilitating rapid, high-throughput processing. Unlike old-fashioned, multi-step RNA isolation protocols, no heating, washing, or centrifugation steps are required. The kit greatly simplifies a laborious 30–60 minute process and reduces it to 7 minutes.

The TaqMan® Gene Expression Cells-to-CT™ Kit workflow enables unsurpassed gene expression evaluation with any of the >700,000 TaqMan® Gene Expression Assays. This kit has been extensively tested for specificity with a broad selection of TaqMan® Gene Expression Assays and shows performance equivalent to that obtained with purified RNA (see figure).

Achieve unsurpassed performance and sensitivity
Unlike some competitor kits that limit the amount of lysate in the RT reaction to 5%, the TaqMan® Gene Expression Cells-to-CT™ Kit can accommodate 45% of the total RT reaction volume as cell lysate. Additionally, cDNA can comprise up to 45% of the real-time PCR reaction volume. The large lysate volume in the optimized RT reaction, along with the large cDNA volume in the subsequent real-time PCR using the TaqMan® Gene Expression Master Mix, lead to maximum sensitivity. The master mix amplifies the target precisely and accurately, enabling the detection of small quantities of target, such as transcripts expressed at low levels.

The ability to detect limited target quantities was tested by mixing a constant number of human cells with various numbers of mouse cells. The cell mixtures were prepared using the TaqMan® Gene Expression Cells-to-CT™ and assayed for a mouse-specific and human-specific gene. Comparative data were generated in the same manner with RNA purified by traditional methodology. The data show that RNA from as few as 10 mouse cells was detectable in a background of RNA from 10,000 human cells (see figure). The ability of the TaqMan® Gene Expression Cells-to-CT™ Kit to detect relative low abundance transcripts was equivalent to that of purified RNA; at low levels, it was superior.

Additionally, the performance of the TaqMan® Gene Expression Cells-to-CT™ Kit was compared to competitor lysate kits and to purified RNA. Inputs of 100–100,000 cells⁄lysis reaction were examined. The sensitivity of the TaqMan® Cells-to-CT™ Kit protocol was equivalent to that obtained with purified RNA, and it surpassed competitor lysates (see figure). Furthermore, the lack of inhibition at high cellular inputs and the low variation among technical replicates demonstrate the reliability of this approach for gene expression studies using cultured cells.

Proven performance, proven together
All components of the TaqMan® Gene Expression Cells-to-CT™ Kit have been optimized for consistent and reliable performance. This removes the guesswork involved in assembling separate sample preparation, RT, and real-time PCR kits. And the TaqMan® Gene Expression Cells-to-CT™ Kit has been validated with TaqMan® Gene Expression Assays for added quality assurance.

Accessory product
The TaqMan® Cells-to-CT™ Control Kit was designed for use with the TaqMan® Gene Expression Cells-to-CT™ Kit. This kit contains an endogenous control (ACTB) to normalize for sample loading variability and an artifical XenoRNA™ control and corresponding TaqMan® Gene Expression Assay to monitor the effects of PCR inhibition.

Computer RAM Upgrade Kit, laptop D600/D505 Applied Biosystems™

This upgrade kit is intended for use with the 7300⁄7500⁄7500 Fast Sequence Detection Systems

For Research Use Only. Not for use in diagnostics procedures.

VetMAX™ M. paratuberculosis 2.0 Kit Thermo Scientific™

Note: The name of this product has changed. It was previously called LSI VetMAX M. paratuberculosis Advanced.

Same as TaqVet M. paratuberculosis Advanced Real-Time PCR Kit

TaqMan® probes - DNA - duplex - One well reaction

Real-time PCR enables sensitive and specific detection of pathogen nucleic acid in animal samples, allowing for reliable and rapid screening and detection of infected animals.

The LSI VetMAX™ kits include:

• 1 ready-to-use mix that includes a set of oligonucleotides for detection of the target gene OR these separate components that will need to be mixed together: a set of oligonucleotides for the detection of the target gene + a TaqMan® real-time PCR master mix + an RT enzyme (only for RNA kits).
• 1 external positive control of the same nature as the target (inactivated bacteria or virus). Use as a quality control for PCR and/or extraction yield.
• 1 internal positive control (exogenous) to be added at the time of extraction. Use as a quality control for extraction yield.
• A quality control certificate.

For veterinary use only.

Regulatory requirements vary by country; products may not be available in your geographic area.

Biological hazard safety

WARNING! BIOHAZARD. Biological samples such as tissues, body fluids, infectious agents, and blood of humans and other animals have the potential to transmit infectious diseases. All work should be conducted in properly equipped facilities using the appropriate safety equipment (for example, physical containment devices). Safety equipment also may include items for personal protection, such as gloves, coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, or goggles. Individuals should be trained according to applicable regulatory and company/institution requirements before working with potentially biohazardous materials. Follow all applicable local, state/provincial, and/or national regulations. The following references provide general guidelines when handling biological samples in laboratory environment.

U.S. Department of Health and Human Services, Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th Edition, HHS Publication No. (CDC) 21-1112, Revised December 2009

World Health Organization, Laboratory Biosafety Manual, 3rd Edition, WHO/CDS/CSR/LYO/2004.11

Purification , Cartridge , 50 nmol Invitrogen™

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3' and 5' modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE).

TaqMan™ Exogenous Internal Positive Control Reagents Applied Biosystems™

Applied Biosystems® TaqMan® Exogenous Internal Positive Control Reagents contain a pre-optimized internal positive control (IPC) with pre-designed primers and TaqMan® probe. The IPC can be spiked into samples to distinguish true target negatives from PCR inhibition. This TaqMan® Exogenous Internal Positive Control Reagents kit allows you to amplify a low-copy target DNA in the same tube with the IPC. Although the target and IPC DNAs may differ in initial copy number, the concentration of the IPC primers in the PCR reaction is limiting so that the amplification efficiency of the target reaction is not compromised.

• Pre-designed primers and TaqMan® probe eliminate assay design

• Rapid assay development guidelines can minimize optimization time

• Separate components provide flexibility in assay set-up

Note: This is not a quantitative kit and should not be used for normalization. Instead, this kit is designed to distinguish types of negative results: a negative call for the target sequence and a positive call for the IPC indicate that no target sequence is present, and a negative call for the target sequence and negative call for the IPC suggest PCR inhibition.

Comprehensive Guidelines Included
Applied Biosystems® has developed a comprehensive set of guidelines to ensure success when using Applied Biosystems® Real-Time PCR reagents and instrumentation. These guidelines are simple and easy to follow. Many traditional variables, such as magnesium chloride concentration and the thermal cycling protocol itself, have been standardized, greatly reducing assay development time.

Normalization Built-In
All Applied Biosystems® Real-Time PCR reagent kits contain a passive internal reference to normalize non-PCR related fluorescence fluctuations. Normalizing with a passive internal reference minimizes well-to-well variability that can result from a variety of causes, such as pipetting errors and sample evaporation. The passive reference is also essential for accurate results when multiple probes (with different reporter dyes) are combined in a single tube.

Note: See user's manual or package insert for limited label license, and trademark information. For Research Use Only. Not for use in diagnostics procedures.

TaqMan™ Fast Cells-to-CT™ Kit Invitrogen™

The TaqMan® Fast Cells-to-CT™ Kit makes it possible to use Fast-cycling, real-time PCR chemistry and instrumentation to perform gene expression analysis directly from cultured cells without RNA purification.

The TaqMan® Fast Cells-to-CT Kit is:

• Complete—optimized workflow includes cell lysis reagents with gDNA removal, RT enzyme mix, RT buffer, and TaqMan® Fast Universal PCR Master Mix
• Fast—prepare samples at room temperature in 7 minutes, including DNase treatment; TaqMan® Fast Universal PCR Master Mix delivers real-time PCR results in ~35 minutes
• Convenient—lyse samples in a tube or directly in 96- or 384-well culture plates
• Robust—perform gene expression analysis on 10 to 100,000 cells per sample, with results equivalent to those from purified RNA
• Efficient—contains sufficient reagents to generate 500 real-time PCR results from 100 starting samples

Complete, validated solution
TaqMan® Cells-to-CT™ technology features a unique method for lysing cultured cells while removing genomic DNA (gDNA) and preserving RNA integrity. Additionally, the kit contains reverse transcription (RT) reagents and TaqMan® Fast Universal PCR Master Mix for Fast real-time PCR analysis, and has been extensively tested and validated with gold-standard TaqMan® Gene Expression assays, sold separately. All kit components have been optimized to work together for consistent and reliable performance, removing the guesswork involved in assembling separate kits for sample preparation, reverse transcription, and Fast real-time PCR.

Fast, convenient workflow
The TaqMan® Fast Cells-to-CT™ Kit incorporates a simple 7-minute sample preparation procedure (see figure) for use with 10 to 100,000 cultured cells directly in 96- or 384-well culture plates or in tubes. Cells are washed in PBS and lysed for 5 minutes at room temperature; an optional DNase treatment can be performed concurrently. Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution. The kit greatly simplifies a traditionally time-consuming, labor intensive process and reduces it to 7 minutes.

The lysates are now ready for reverse transcription with the optimized RT system included in the kit, or they can be stored at -20°C for up to 5 months, for future analysis. The supplied TaqMan® Fast Universal PCR Master Mix delivers sensitive and reproducible results approximately 3 times faster than standard PCR reagents and instruments. The combination of fast and simple sample preparation, and Fast-cycling real-time PCR results (~35 minutes) enables more rapid quantitation of gene targets than previously available. The TaqMan® Fast Cells-to-CT™ Kit workflow reduces the time from sample to result by 40% or more, compared to traditional RNA purification and standard real-time PCR.

Robust, sensitive, and reliable results
Accurate gene expression analysis over a wide range of sample input for genes with various expression levels has traditionally been a challenge for researchers. Therefore, a robust solution is important for profiling RNA from a few or many cells. Shown is the demonstration of the excellent signal linearity (R2 = 0.997) observed using lysates from TaqMan® Fast Cells-to-CT™ Kit across 5 logs of cellular input (10–105 cells per lysis reaction) (see figure). Thus, a single kit can be used for analysis of gene expression from limited or dense samples, with no compromise in data integrity caused by PCR inhibitors.

PCR performance of cell lysates using the TaqMan® Fast Cells-to-CT™ Kit was equivalent to that from purified RNA, using the supplied TaqMan® Fast Universal PCR Master Mix (see figure). Both TaqMan® Cells-to-CT™ Kit lysates and purified RNA showed linear detection over a 5 log range of cell input equivalents. The enhanced sensitivity at the 10-cell input with TaqMan® Fast Cells-to-CT™ Kit lysates compared to purified RNA, using the same RT and PCR reagents, results from more efficient retention of target molecules during preparation, i.e., no loss of RNA typically associated with sample transfer or heating.

The TaqMan® Fast Cells-to-CT™ Kit also demonstrated superior performance compared to purified RNA which was reverse transcribed and PCR amplified with Competitor Q’s RT and Fast Master Mix. The observed dynamic range using Competitor Q’s reagents with Fast cycling was 102 to 105 cell equivalents, or 1 log less than TaqMan® Cells-to-CT™ Kit lysates or purified RNA amplified with TaqMan® Fast Universal PCR Master Mix (see figure). The TaqMan® Fast Universal PCR Master Mix is formulated to deliver sensitive and reproducible results faster than is possible with standard reagents and instruments, and is robust across a wide range of target genes and concentrations.

Custom Primer, 3' RHD Invitrogen™

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3' and 5' modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE).

Computer RAM Upgrade Kit, laptop D510/D520 Applied Biosystems™

This upgrade kit is intended for use with the 7300⁄7500⁄7500 Fast Sequence Detection Systems.

For Research Use Only. Not for use in diagnostics procedures.

5' Mod , Pacific Blue Dye , 200 nmol Invitrogen™

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3' and 5' modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE).

QuantStudio™ 3D Digital PCR System Package, with Master Mix, Chip Kit v2, and customer site training Applied Biosystems™

Bring digital PCR into your lab with a QuantStudio™ 3D Digital PCR System package. This version includes everything required to start a digital PCR experiment:

• QuantStudio™ 3D Digital PCR Instrument
• QuantStudio™ 3D Digital PCR Chip Loader
• ProFlex™ 2 x Flat Block Thermal Cycler
• 96 QuantStudio™ 3D Digital PCR Chips v2
• QuantStudio™ 3D Digital PCR Master Mix v2 for 96 chips
• Warranty Service (1 year)
• 1-Day Field Applications Scientist-lead Training at Customer Site

In addition to the above, this package includes additional reagents and consumables for use during the Field Applications Scientist-lead Training:
• QuantStudio™ 3D Digital PCR Starter Kit (includes DNA, assay, chips and master mix)

>> Download QuantStudio™ 3D Digital PCR Software v3.0  |  [User Instructions]

QuantStudio 3D Digital PCR System
The Applied Biosystems™ QuantStudio™ 3D Digital PCR System is an affordable digital PCR platform with high accuracy, sensitivity, and precision in a scalable small-footprint platform that is simple to use at any experience level.

• Simple load and go, turn-key chip-based workflow
• Save money with a system priced at less than 50% of droplet-based digital PCR platforms
• Quantitate absolute nucleic acid concentrations in copies/µL

Digital PCR makes new applications possible by enabling sensitive and precise absolute target quantification without the use of a reference or standard curve. Researchers can now go beyond Ct to absolute quantification for investigations that involve detecting extremely rare events or determining the exact number of targets in a sample. The simple, affordable, and contained QuantStudio 3D Digital PCR System brings digital PCR to all researchers.

The QuantStudio 3D Digital PCR System leverages a high-density nanofluidic chip technology to partition a sample into as many as 20,000 independent reaction wells. Samples and amplification products remain completely contained throughout the streamlined workflow, and an easily interpretable answer (target copies/µL) is produced—all of which minimizes the level of expertise required to quantify specific nucleic acid sequences of interest. Simply load the reaction mix onto the chip, amplify on the ProFlex™ 2 x Flat Block Thermal Cycler and read the target concentration on the QuantStudio 3D Digital PCR Instrument.

Custom Primer Invitrogen™

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3' and 5' modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE).

Custom Primer Invitrogen™

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3' and 5' modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE).

Oligo (dT) Primer (50 µM) Invitrogen™

These are the same Ambion® primers that are currently included in the RETROscript® Kit (SKU# AM1710). They are provided at a stock concentration of 50 µM and are functionally tested using the RETROscript® Kit.
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