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AmpliTaq Gold™ 360 DNA Polymerase Applied Biosystems™

The Applied Biosystems® AmpliTaq Gold® 360 DNA Polymerase, when used with improved AmpliTaq Gold 360 Buffer, and the optional 360 GC Enhancer, amplifies a vast range of DNA sequence contexts. AmpliTaq Gold® 360 DNA Polymerase delivers 360° coverage for a full range of targets.
* Optimized for the broadest range of targets—from everyday to challenging
* Unmatched sensitivity, specificity, and yield
* Robust amplification of GC-rich sequences with market-leading 360 GC Enhancer
* Achieves the highest quality sequencing data

Optimized for Easy and Challenging Targets

Challenging targets include AT-rich, GC-rich, primer-dimer forming amplicons, homopolymer repeats, and amplicons that pose sequencing challenges. Amplicons that previously required specialized enzymes and reaction conditions can now be reproducibly amplified with a single reagent under standardized conditions (Figure 1). Competitive benchmarking across more than 40 amplicons distinguishes AmpliTaq Gold® 360 as the best-performing enzyme, ensuring the highest probability of success for the amplification of both everyday and challenging targets (Table 1).
As shown in Figure 1, GC-rich regions are especially poorly amplified with competitor DNA polymerases and the original AmpliTaq Gold, while AmpliTaq Gold 360 provides successful, robust amplification.

Optimized for Hot-Start PCR

AmpliTaq Gold 360 DNA Polymerase provides the same hot-start specificity as AmpliTaq Gold DNA polymerase. A high-temperature incubation step is required to activate AmpliTaq Gold DNA Polymerase, which ensures that the active enzyme is generated only at temperatures in which the DNA is fully denatured and when the primers are not annealed.
When the polymerase is added to the reaction mixture at room temperature, primer extension does not occur because the enzyme is inactivated. Any low stringency mispriming events that may have occurred will not be enzymatically extended and will not be amplified. Hence, PCR setup can be performed at room temperature without concern for extension at misprimed sites. The amount of AmpliTaq Gold 360 DNA polymerase increases in the reaction slowly with each cycle number, and specific product yield increases without buildup of nonspecific products, including primer dimers. Excellent specificity across a broad range of targets is shown in Figure 1 and is summarized in Figure 2. The exquisite specificity allows easier multiplexing and allelic discrimination.
The amount of AmpliTaq Gold 360 DNA polymerase increases in the reaction slowly with each cycle number, and specific product yield increases without buildup of nonspecific products, including primer dimers. Excellent specificity across a broad range of targets is shown in Figure 1 and is summarized in Figures 2A & B. The exquisite specificity allows easier multiplexing and allelic discrimination.

Superior Sensitivity and Amplification Length

Compared to the original AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process to eliminate contaminating bacterial DNA sequences from the enzyme preparation. When used with the enhanced 360 Gold Buffer, this ultra-pure enzyme, in addition to its hot-start capabilities, reduces false positive results, amplifies low-level target sequences, and promotes the amplification of a variety of templates including those from bacterial and human genomes.
AmpliTaq Gold 360 DNA Polymerase efficiently amplifies targets present at low copy number (Figure 3), even in the presence of high concentrations of complex DNA, making it especially suited for low-copy pathogen detection, and amplification of targets from degraded DNA samples. The extreme purity of the enzyme contributes to its unmatched sensitivity. AmpliTaq Gold 360 DNA Polymerase efficiently and reproducibly amplifies long (up to 5 Kb) sequences. Figure 4 demonstrates robust PCR amplification of long human and plasmid DNA.
 

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.

AmpliTaq™ 360 DNA Polymerase Applied Biosystems™

The Applied Biosystems AmpliTaq360 DNA Polymerase, when used with the new enhanced AmpliTaq 360 Buffer and the optional 360 GC Enhancer, amplifies a vast range of DNA sequence contexts. Compared to the original AmpliTaq DNA polymerase, AmpliTaq 360 polymerase is purified by an additional proprietary separation process which reduces contaminating bacterial DNA sequences from the enzyme preparation. This ultra-pure enzyme reduces false positive results, amplifies low-level target sequences, and when combined with the proprietary AmpliTaq 360 Buffer Kit, promotes the amplification of a variety of templates including those from bacterial and human genomes.

• Optimized for the broadest range of targets—from everyday to challenging
• Unmatched sensitivity and yield
• Market-leading GC enhancer for robust amplification of GC-rich sequences
• Achieves the highest quality sequencing data

For superior PCR performance, we recommend DreamTaq DNA Polymerase.

TaqMan™ RNase P Instrument Verification Plate for ViiA™ 7 System, 384-well Applied Biosystems™

Used to perform verification of your ViiA™7 System with 384 Well Block. Please see your instrument maintenance guide for full instructions on use.

M13 Human, Reverse Primer, HPLC Applied Biosystems™

Use our tool to find and order pre-designed primers for PCR and Sanger sequencing. Free of SNPs and dimers, target-specific, and suitable for universal PCR conditions, these primers can be ordered unmodified, M13-tailed, HPLC-purified, or desalted. All primers are checked by mass spectrometry.

QuantStudio™ 6 Pro Real-Time PCR System, 96-well, 0.2 mL, desktop Applied Biosystems™

The Applied Biosystems QuantStudio 6 Pro Real-Time PCR System adds innovative smart features to the real-time PCR (qPCR) workflow, all in a compact footprint. With facial authentication, voice commands, Smart Help, multiplex capability, motorized interchangeable blocks, simplified workflow, intuitive software, and large capacitive touch-screen interface, the QuantStudio 6 Pro system offers exceptional reproducibility with minimal well-to-well and instrument-to-instrument variation. Designed for ultimate ease-of-use, the system offers high quality, excellent reliability, and an optimal user experience for researchers who want to work smarter in the lab.

Features of the QuantStudio 6 Pro Real-Time PCR System include:
• Easy and convenient hands-free operation with facial authentication and voice commands
• Reduced errors and manual entries with smart qPCR workflow and TaqMan Array Plates with RFID
• Space savings with small footprint and true standalone operation; set-up, start, monitor, and analyze directly on the touchscreen
• Peace of mind with Smart Help; contact Technical Support directly from the touchscreen
• Reduced downtime with Smart Remote Support; show issues to Technical Support for speedy resolution
• Application versatility with easily changed blocks and three different block types
• Access, analyze, and share data anytime, anywhere with our Connect cloud-based platform for real-time information access, secure data storage, and online results sharing
• Remotely monitor experiments using the Instrument Connect monitoring app
• Design and Analysis software compatible with PC and Mac operating systems
• Compatible with over 8 million TaqMan Assays for gene expression, genetic variation, gene regulation, or protein expression experiments

Easy and convenient hands-free operation
The QuantStudio 6 Pro system was designed for a simple and powerful qPCR experience. Hands-free operation with facial authentication and voice commands enables users to quickly and easily access their personal profiles and settings without even removing their gloves.

Smart qPCR for ultimate ease of use
With embedded RFID readers, the QuantStudio 6 Pro system has the capability to run TaqMan Array Plates with RFID tags. These RFID tags simplify data tracking (with embedded lot number, expiration date, etc.) and reduce the need for manual data entry with preloaded assay layout, dye information, and thermal protocol. Automating this data transfer allows users to simply load the plate and start the run—no data entry needed.

Learn more about the TaqMan Array Plates with RFID ›

Peace of mind with Smart service
Minimize downtime with premium service offerings for the QuantStudio 6 Pro system. Smart Help allows users to contact Technical Support directly from the touchscreen, without the need to download files or spend time on the phone. Experienced technicians will analyze the request and respond according to the user’s preference (phone or email) and resolve the issue as quickly as possible.

Smart Remote Support is an immersive collaboration tool that allows customers to quickly and efficiently show issues to Technical Support for resolution, helping reduce instrument downtime compared to on-site repair and helping minimize the need for on-site visits.

Connect
The Connect platform provides scientists with a secure place to store, analyze, and share data. Applied Biosystems qPCR Analysis Modules within the platform are web browser-based software tools that are flexible, fast, and easy to use. These software tools support a variety of analyses including relative quantitation, standard curve, high resolution melt (HRM), genotyping, and presence/absence. They are compatible with Mac or PC as well as most Applied Biosystems instrument file types.

Format flexibility
The QuantStudio 6 Pro system accommodates the interchange of 96-well (0.2 mL), 96-well (0.1 mL), and 384-well blocks. The system is delivered with one block type; other block types may be purchased separately. For a system that also supports TaqMan array cards (TAC), as well as automation, please see our QuantStudio 7 Pro Real-Time PCR System. The QuantStudio 6 Pro system can also be upgraded to a QuantStudio 7 Pro system at a later date, as your needs change. And for the ultimate in flexibility, consider the QuantStudio 12K Flex Real-Time PCR System, which also accommodates TaqMan OpenArray plates.

Upgrade capabilities when you need them
The QuantStudio 6 Pro system can be seamlessly upgraded to a QuantStudio 7 Pro system by a field service engineer to give you additional automation, throughput, and multiplexing capabilities.

High application versatility
Simple workflows, reliable assays and reagents, multiple formats, and intuitive software are available for a broad range of applications, including gene expression, genetic variation, and gene regulation.

Results you can trust
Improved well-to-well and instrument-to-instrument data accuracy is achieved with the OptiFlex system, which features five coupled excitation and emission filter channels.

Sensitive data analysis
Detect changes in target quantity as small as 1.5-fold in single-plex reactions and obtain 10 logs of linear dynamic range.

AmpliTaq™ 360 Buffer, 25 mM MgCl2 and 360 GC Enhancer Applied Biosystems™

The Applied Biosystems® AmpliTaq® 360 Buffer Kit is used in conjection with the AmpliTaq 360 DNA polymerase for those few experiments when the reagents supplied with the enzyme are not sufficient for the researcher's needs. When used together with or without the optional 360 GC Enhancer, amplifies a vast range of DNA sequence contexts. AmpliTaq® 360 DNA Polymerase delivers 360° coverage for a full range of targets.

* Optimized for the broadest range of targets—from everyday to challenging
* Robust amplification of GC-rich sequences with market-leading 360 GC Enhancer
* Achieves the highest quality sequencing data

Optimized for Easy and Challenging Targets
Challenging targets include AT-rich, GC-rich, primer-dimer forming amplicons, homopolymer repeats, and amplicons that pose sequencing challenges. Amplicons that previously required specialized enzymes and reaction conditions can now be reproducibly amplified with a single reagent under standardized conditions (Figure 1). Competitive benchmarking across more than 40 amplicons distinguishes AmpliTaq Gold® 360 as the best-performing enzyme, ensuring the highest probability of success for the amplification of both everyday and challenging targets (Table 1).

As shown in Figure 1, GC-rich regions are especially poorly amplified with competitor DNA polymerases and the original AmpliTaq Gold, while AmpliTaq Gold 360 provides successful, robust amplification.

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.

M13 Sequencing Forward Primer, HPLC Applied Biosystems™

Use our tool to find and order pre-designed primers for PCR and Sanger sequencing. Free of SNPs and dimers, target-specific, and suitable for universal PCR conditions, these primers can be ordered unmodified, M13-tailed, HPLC-purified, or desalted. All primers are checked by mass spectrometry.

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.

Axiom™ Cotton Genotyping Array Applied Biosystems™

Axiom Cotton Genotyping Array was designed through Affymetrix- Expert Design Program, and the markers on the array were selected from public databases in collaboration with the National Botanical Research Institute, India. The array includes 35,550 markers that were discovered in G. hirsutum and G. barbadense.

Axiom Cotton Genotyping Array can be used to analyze samples from different populations by adding up to 380,000 custom markers or by transferring polymorphic markers with 100%; fidelity on to the Axiom 384HT myDesign™ breeders array. The Axiom GT1 algorithm can automatically cluster, assign genotypes, and classify the markers into six different categories for easy visualization.

Features of Axiom Cotton Genotyping Array
Comprehensive content:
The Axiom Cotton genotyping array includes a total of 35,550 markers:
• 28,158 intra-specific markers identified using gene-enriched genomic sequences of G. hirsutum.
• 7,392 markers discovered using genome reduction methodology that is based on restriction site conservation (GR-RSC).
   - 5,286 markers were discovered in an inter-specific assembly of G. hirsutum and G. barbadense.
   - 2,106 markers are intra-specific to G. hirsutum.

Applications of Axiom Cotton Genotyping Array
Complex trait research and molecular breeding:
• GWAS and QTL mapping
• Identification of economically-important traits
• Improved accuracy marker-assisted selection programs through genomic selection

Required Products
GeneChip Command Console Software
GeneTitan Multi-Channel Instrument

QuantStudio™ 6 Pro Real-Time PCR System, 96-well, 0.1 mL, desktop Applied Biosystems™

The Applied Biosystems QuantStudio 6 Pro Real-Time PCR System adds innovative smart features to the real-time PCR (qPCR) workflow, all in a compact footprint. With facial authentication, voice commands, Smart Help, multiplex capability, motorized interchangeable blocks, simplified workflow, intuitive software, and large capacitive touch-screen interface, the QuantStudio 6 Pro system offers exceptional reproducibility with minimal well-to-well and instrument-to-instrument variation. Designed for ultimate ease-of-use, the system offers high quality, excellent reliability, and an optimal user experience for researchers who want to work smarter in the lab.

Features of the QuantStudio 6 Pro Real-Time PCR System include:
• Easy and convenient hands-free operation with facial authentication and voice commands
• Reduced errors and manual entries with smart qPCR workflow and TaqMan Array Plates with RFID
• Space savings with small footprint and true standalone operation; set-up, start, monitor, and analyze directly on the touchscreen
• Peace of mind with Smart Help; contact Technical Support directly from the touchscreen
• Reduced downtime with Smart Remote Support; show issues to Technical Support for speedy resolution
• Application versatility with easily changed blocks and three different block types
• Access, analyze, and share data anytime, anywhere with our Connect cloud-based platform for real-time information access, secure data storage, and online results sharing
• Remotely monitor experiments using the Instrument Connect monitoring app
• Design and Analysis software compatible with PC and Mac operating systems
• Compatible with over 8 million TaqMan Assays for gene expression, genetic variation, gene regulation, or protein expression experiments

Easy and convenient hands-free operation
The QuantStudio 6 Pro system was designed for a simple and powerful qPCR experience. Hands-free operation with facial authentication and voice commands enables users to quickly and easily access their personal profiles and settings without even removing their gloves.

Smart qPCR for ultimate ease of use
With embedded RFID readers, the QuantStudio 6 Pro system has the capability to run TaqMan Array Plates with RFID tags. These RFID tags simplify data tracking (with embedded lot number, expiration date, etc.) and reduce the need for manual data entry with preloaded assay layout, dye information, and thermal protocol. Automating this data transfer allows users to simply load the plate and start the run—no data entry needed.

Learn more about the TaqMan Array Plates with RFID ›

Peace of mind with Smart service
Minimize downtime with premium service offerings for the QuantStudio 6 Pro system. Smart Help allows users to contact Technical Support directly from the touchscreen, without the need to download files or spend time on the phone. Experienced technicians will analyze the request and respond according to the user’s preference (phone or email) and resolve the issue as quickly as possible.

Smart Remote Support is an immersive collaboration tool that allows customers to quickly and efficiently show issues to Technical Support for resolution, helping reduce instrument downtime compared to on-site repair and helping minimize the need for on-site visits.

Connect
The Connect platform provides scientists with a secure place to store, analyze, and share data. Applied Biosystems qPCR Analysis Modules within the platform are web browser-based software tools that are flexible, fast, and easy to use. These software tools support a variety of analyses including relative quantitation, standard curve, high resolution melt (HRM), genotyping, and presence/absence. They are compatible with Mac or PC as well as most Applied Biosystems instrument file types.

Format flexibility
The QuantStudio 6 Pro system accommodates the interchange of 96-well (0.2 mL), 96-well (0.1 mL), and 384-well blocks. The system is delivered with one block type; other block types may be purchased separately. For a system that also supports TaqMan array cards (TAC), as well as automation, please see our QuantStudio 7 Pro Real-Time PCR System. The QuantStudio 6 Pro system can also be upgraded to a QuantStudio 7 Pro system at a later date, as your needs change. And for the ultimate in flexibility, consider the QuantStudio 12K Flex Real-Time PCR System, which also accommodates TaqMan OpenArray plates.

Upgrade capabilities when you need them
The QuantStudio 6 Pro system can be seamlessly upgraded to a QuantStudio 7 Pro system by a field service engineer to give you additional automation, throughput, and multiplexing capabilities.

High application versatility
Simple workflows, reliable assays and reagents, multiple formats, and intuitive software are available for a broad range of applications, including gene expression, genetic variation, and gene regulation.

Results you can trust
Improved well-to-well and instrument-to-instrument data accuracy is achieved with the OptiFlex system, which features five coupled excitation and emission filter channels.

Sensitive data analysis
Detect changes in target quantity as small as 1.5-fold in single-plex reactions and obtain 10 logs of linear dynamic range.

Pharmaceutical Analytics QuantStudio™ 5 Real-Time PCR System, 384-well, desktop Applied Biosystems™

The Applied Biosystems™ QuantStudio™ 5 Real-Time PCR System for Pharmaceutical Analytics is designed for users who need superior performance, maximum dye versatility, and security options in a real-time PCR system that is affordable and easy to use. The QuantStudio 5 pharmaceutical analytics system is used together with specialized sample preparation and quantitation kits for host cell protein, host cell DNA, and Protein A impurity testing of biological products produced in cell culture.The optimized Design and Analysis software is ideal for both first-time and experienced users. When connected to the Thermo Fisher Cloud, the QuantStudio 5 system provides access to your data wherever and whenever you want. Using proven OptiFlex® technology (featuring 6 decoupled channels and white LED) and featuring six independent Veriflex® temperature zones, the QuantStudio 5 96-well system enables improved data accuracy and sensitivity for a broad range of genomic applications. The QuantStudio 5 system also offers the built-in software features of electronic record security and prevention of unauthorized instrument access to assist with 21 CFR Part 11 compliance.

Features of the QuantStudio 5 system with 96-well block include:

• Proven OptiFlex technology (6 decoupled channels/21 filter combinations with white LED) for trusted results
• User account management, locked workflow, and Pause features for greater control of experiment data
• Built-in software features to support compliance with 21 CFR Part 11 guidelines
• Secure cloud-based software provides increased accessibility and storage options
• Simplified instrument software with interactive touchscreen for greater ease of use
• 96-well 0.2mL format or 96-well 0.1mL format

Thermo Fisher Cloud
Thermo Fisher Cloud provides scientists with a secure place to store, analyze, and share data. Upload data from the instrument or upload your own files from anywhere you have web access. Applied Biosystems qPCR Analysis Modules within the Thermo Fisher Cloud are web browser-based software tools that are flexible, fast and easy to use.

Downloads
>> QuantStudio 3/5 Design & Analysis Software | Release Notes | Example Data & Templates
>> QuantStudio 3/5 Firmware | Release Notes

Easy to use
The QuantStudio 5 Real-Time PCR System is compact in form, factory calibrated and is delivered in user-friendly packaging to enable fast and simple installation for immediate use. The interactive touchscreen provides a modern usability experience and the simplified instrument software/pre-optimized protocol templates allow users of any experience level to quickly set up and start a run.

Design and analysis software options:

Option 1: QuantStudio Design and Analysis cloud software
• Supports web browser-based system configuration with PC or MAC computer
• Streamlined software for improved usability and response time
• Enables secure access of your data when and where you want through the cloud
• No software to install, no licenses and no versions to update

Option 2: QuantStudio Design and Analysis desktop software
• Supports traditional co-located computer system configuration on Windows® 7 OS
• Streamlined software for improved usability and response time
• No licenses to purchase

Total control
With built-in software features to support compliance with 21 CFR Part 11 guidelines, user account management, locked workflow and Pause features, the QuantStudio 5 system offers users greater control of experiment data

Multiplex performance results that you can trust
Improved well-to-well and instrument-to-instrument data accuracy is achieved with OptiFlex technology. In the 96-well system this technology features 6 decoupled excitation and emission filter channels, for a total of 21 filter combinations, with white LED.

Sensitive data analysis
Detect changes in target quantity as small as 1.5-fold in single-plex reactions and obtain 10 logs of linear dynamic range.

High application versatility
Simple and optimized protocols, reliable assays and reagents, and intuitive software are available for a broad range of applications, including gene expression, genetic variation, gene regulation, and more.

qualyfast™ Legionella PCR and DNA Extraction Kits Thermo Scientific™

Detect both Legionella species and Legionella pneumophila from water samples with this rapid, sensitive and specific solution enabled by the qualyfast® Legionella PCR Assay, Free DNA Inactivator, DNA Extraction and Quantification Standards kits.The assay independently detects both Legionella species and L. pneumophilia, includes inhibition control to minimize false-negative results, and offers the option for quantification by using calibrated standards. All consumables required for the workflow are included within the kits, with lyophilized and pre-dosed PCR reagents for ease-of-use.

AmpliTaq Gold™ 360 Buffer Kit Applied Biosystems™

The AmpliTaq Gold® 360 Buffer Kit is used in conjunction with AmpliTaq Gold® 360 DNA Polymerase and is intended for those occasions when the quantity of reagents supplied with the enzyme are not sufficient. The kit includes 1.5 mL (each) of 10X AmpliTaq Gold 360 Buffer, 25 mM MgCl2, and 3360 GC Enhancer. Typically, 1.5 mL is sufficient for use with 250 U of enzyme.

M13 Sequencing Reverse Primer, HPLC Applied Biosystems™

Use our tool to find and order pre-designed primers for PCR and Sanger sequencing. Free of SNPs and dimers, target-specific, and suitable for universal PCR conditions, these primers can be ordered unmodified, M13-tailed, HPLC-purified, or desalted. All primers are checked by mass spectrometry.

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.
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