Product Class: Other

WarmStart® RTx Reverse Transcriptase (Glycerol-free)

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Catalog #M0439

Product Introduction

  • Glycerol-free version of WarmStart RTx Reverse Transcriptase (NEB #M0380) to support lyophilization and automation workflows
  • Ideal for use in Loop-mediated Isothermal Amplification (LAMP)
  • WarmStart® technology enables high-throughput applications, room temperature reaction setup, and increases the consistency and specificity of amplification reactions
  • Sensitive RNA detection: 1 pg RNA in LAMP, 10 fg input RNA in RT-PCR
  • Optimal reaction temperature from 55-65°C
  • One of several products available glycerol-free

Product Information

Description

WarmStart RTx Reverse Transcriptase (Glycerol-free) is formulated without glycerol to support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows.

WarmStart RTx Reverse Transcriptase is a unique in silico-designed RNA-directed DNA polymerase coupled with a reversibly-bound aptamer that inhibits RTx activity below 40°C. This enzyme can synthesize a complementary DNA strand initiating from a primer using RNA (cDNA synthesis) or single-stranded DNA as a template. RTx is a robust enzyme for RNA detection in amplification reactions and is particularly well suited for use in LAMP (Loop-mediated Isothermal Amplification). The WarmStart property enables high throughput applications, room temperature setup, and increases the consistency and specificity of amplification reactions. RTx contains intact RNase H activity.

WarmStart RTx Reverse Transcriptase (Glycerol-free) offers the same robust detection of human RNA targets in RT-LAMP assays as the glycerol-containing enzyme.

M0439 Targets

RT-LAMP (RNA targets) experiments were performed with NEB #M0380: WarmStart RTx Reverse Transcriptase and NEB #M0538: Bst 2.0 WarmStart DNA Polymerase, or NEB #M0439: WarmStart RTx Reverse Transcriptase (Glycerol-free) and NEB #M0402: Bst 2.0 WarmStart DNA Polymerase (Glycerol-free). Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent dye were set up in quadruplicate over three logs of total Jurkat RNA (10 ng to 0.1 ng) in 96-well, 25 µl reactions. Control reactions without template (NTC) were also evaluated. Reactions were incubated at 65°C for 40 minutes and fluorescence was monitored every 15 seconds in the SYBR/FAM channel of a real-time thermocycler (Bio-Rad® CFX96). Each dot represents the time at which the fluorescence signal for a single reaction crosses the instrument-defined threshold. All four replicates were detected at each template input unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). Overall, similar performance was observed for both glycerol-containing (NEB #M0538 and NEB #M0380) and glycerol-free (NEB #M0402: and NEB #M0439) enzymes at each template input. No amplification was observed in any of the no template control reactions.

This product is related to the following categories:
Isothermal Amplification & Strand Displacement Products,
cDNA Synthesis & Reverse Transcriptases Products,
PCR, qPCR & Amplification Technologies Products,
This product can be used in the following applications:
Loop-Mediated Isothermal Amplification,
DNA Amplification, PCR & qPCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0439L     -80    
      WarmStart® RTx Reverse Transcriptase (Glycerol-free) M0439LVIAL -80 1 x 0.027 ml 75,000 units/ml
      10X Isothermal Amplification Buffer (Lyo-compatible) B1714SVIAL -20 1 x 1.25 ml 10 X

Properties & Usage

Protocols, Manuals & Usage

Protocols

  1. Typical RT-LAMP Protocol (NEB #M0439)
  2. Typical cDNA Synthesis Protocol with WarmStart® RTx Reverse Transcriptase (Glycerol-free) (NEB #M0439)

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is LAMP and RT-LAMP?
  2. What advantages does WarmStart® RTx provide?
  3. How sensitive is RNA amplification with WarmStart® RTx?
  4. Can WarmStart® RTx be inactivated?
  5. Is WarmStart® RTx active in other buffers?
  6. What is the maximum length of cDNA product produced by WarmStart® RTx?
  7. Does WarmStart® RTx have RNase H activity?
  8. Can DNA be used as a template for WarmStart® RTx?
  9. What are Hot Start and WarmStart® polymerases and when would I use them?
  10. What is the difference between buffers B1714: 10X Isothermal Amplification Buffer (Lyo-compatible) and B0537: 10X Isothermal Amplification Buffer?
  11. What is the Mg2+ concentration in the 10X Isothermal Amplification Buffer (Lyo-compatible)?
  12. Does B1714: 10X Isothermal Amplification Buffer (Lyo-compatible) include any excipients?
  13. Can WarmStart® RTx Reverse Transcriptase (Glycerol-free) at 75,000 U/mL be diluted prior to use?
  14. How many Freeze/Thaw cycles can WarmStart® RTx Reverse Transcriptase (Glycerol-free) tolerate?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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