WarmStart® RTx Reverse Transcriptase

#M0380

cloned at NEB recombinant NEBU 80 Heat
Catalog # Size Concentration Price (£)
M0380L 250 REACTIONS 15,000 units/ml £265.00
M0380S 50 REACTIONS 15,000 units/ml £67.00
  • Ideal for use in Loop-Mediated Isothermal Amplification (LAMP))

  • WarmStart® feature enables high-throughput applications, room temperature reaction setup, and increases the consistency and specificity of amplification reactions




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.

Improved Speed and Sensitivity in RT-LAMPImproved Speed and Sensitivity in RT-LAMP
RT-LAMP reactions with Bst 2.0 WarmStart DNA Polymerase and the indicated reverse transcriptase were incubated at 65°C with 1 pg - 100 ng of Jurkat total RNA. Reactions were monitored with real-time fluorescence, and the resulting curves are shown on left, with the corresponding threshold times on right. WarmStart RTx provides faster reaction threshold times for improved consistancy and sensitivity with lower input RNA amounts. RT-LAMP reactions performed with AMV Reverse Transcriptase resulted in inconsistent detection, as indicated by wide variation at lower RNA input concentrations (blue curves).Sensitive Detection with Reduced False Negatives

Total RNA WarmStart RTx AMV RT

100 ng

3/3

3/3

10 ng

3/3

3/3

1 ng

3/3

3/3

100 pg

3/3

2/3

10 pg

3/3

2/3

1pg

3/3

1/3


RT-LAMP reactions were performed with WarmStart RTx and AMV Reverse Transcriptase with total RNA at 6 input concentrations. Positive detection, indicated by fluorescent detection with a twenty-minute reaction time, was achieved down to 1 pg of total input RNA. Use of AMV resulted in false negatives when total RNA was 100 pg or lower.WarmStart Control of WarmStart RTxWarmStart Control of WarmStart RTx
cDNA synthesis was performed for 10 minutes followed by qPCR analysis. Resulting Cts were normalized against both a "no RT" control for 0% activity and the fastest Ct for 100% activity. WarmStart RTx is inhibited by a reversibly-bound aptamer at temperatures below 40°C.Robust RT-LAMP Across a Range of TargetsRobust RT-LAMP Across a Range of Targets
A variety of RT-LAMP targets were amplified using Bst 2.0 WarmStart DNA Polymerase and the indicated reverse transcriptase. Reactions with WarmStart RTx resulted in faster threshold times and more consistent reaction success across a range of targets and copy number. 

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

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