WarmStart® RTx 反转录酶 | NEB

品牌	货号	产品中文名称	产品英文名称
NEB	M0380	WarmStart® RTx 反转录酶	WarmStart®RTx Reverse Transcriptase

货号	产品中文名称	产品英文名称	规格
M0380L	WarmStart® RTx 反转录酶	WarmStart® RTx Reverse Transcriptase	250 rxns
M0380S	WarmStart® RTx 反转录酶	WarmStart® RTx Reverse Transcriptase	50 rxns

产品特点

• 环介导等温扩增（LAMP）的理想选择
• WarmStart^® 技术有助于在室温下、高通量建立反应体系，提高了扩增反应的一致性和特异性

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.

To support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows, WarmStart RTx Reverse Transcriptase is also offered in a glycerol-free format.

Improved 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 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 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.

产品来源

An E. coli strain that carries the WarmStart RTx Reverse Transcriptase gene.

产品类别:

RT-PCR 产品,

cDNA 合成和反转录酶产品

应用:

Strand Displacement Amplification & Nicking Enzyme,

cDNA Synthesis,

Reverse Transcription (cDNA Synthesis),

Loop-Mediated Isothermal Amplification,

RT-PCR & cDNA Synthesis,

Isothermal Amplification,

PCR

备注：以上仅为 M0380 的部分产品信息，如需NEB货号 M0380 的完整产品信息及相关的实验操作手册等等，请通过NEB官网查找对应的货号获取。