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
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.
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.
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