Sensiscript RT Kit

For reverse transcription of less than 50 ng RNA per reaction for end-point PCR

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Sensiscript RT Kit (200)

Cat no. / ID.   205213

For 200 reverse-transcription reactions: Sensiscript Reverse Transcriptase, 4 x 150 µl 10x Buffer RT, 4 x 100 µl dNTP Mix (contains 5 mM each dNTP), 4 x 1.1 ml RNase-free water
1 269,00 €
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The Sensiscript RT Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Sensitive detection of as few as 10 copies of template
  • High cDNA yields due to high-affinity enzyme
  • No additional RNase H digestion step required
  • Fast and easy procedure with no tedious pipetting steps

Product Details

The Sensiscript Reverse Transcriptase (RT) Kit is ideally suited for highly sensitive applications using very small amounts of RNA (less than 50 ng), such as single-cell RT-PCR and analysis of biopsies and LMD samples. Sensiscript Reverse Transcriptase allows highly efficient RT-PCR over a wide dynamic range and extremely sensitive RT-PCR with very small RNA amounts due to its high affinity for RNA. The Sensiscript RT Kit includes Sensiscript Reverse Transcriptase, dNTPs, and an optimized reaction buffer. Simply add primers for fast and easy cDNA synthesis.

Performance

Sensiscript Reverse Transcriptase (RT) provides outstanding efficiency over a wide dynamic range and high sensitivity for very small amounts of RNA (see figure " RT-PCR with RNA corresponding to 1 cell"). Regions of RNA with high GC content can cause other reverse transcriptases to stop, dissociate from the RNA template, or skip over looped-out regions of RNA (see figure  Full-length RT-PCR products — B). These difficult templates, however, prove no problem for QIAGEN reverse transcriptases (see figure  Full-length RT-PCR products — A). With no need for optimization, the Sensiscript RT Kit makes reverse transcription virtually trouble-free.

Lot-to-lot reproducibility of Sensiscript RT Kits is ensured by rigorous quality control at QIAGEN. The optimized Buffer RT, dNTPs, and water included in all Sensiscript RT Kits are guaranteed RNase-free, and each lot of Sensiscript RT is thoroughly tested for RT-PCR reproducibility.

See figures

Principle

Sensiscript Reverse Transcriptase (RT) is the first commercially available enzyme designed for highly sensitive applications with very small amounts of RNA (<50 ng), such as single-cell RT-PCR or analysis of small biopsies. It is provided ready-to-use with dNTPs and in an optimized reaction buffer that, together with the high affinity of Sensiscript RT for RNA, results in highly specific and sensitive RT-PCR, even with low-copy transcripts, and the ability to read through even complex RNA secondary structures without adjusting temperature or reaction conditions.  Please note that the primer mix is not provided.

For viral RNA, Omniscript RT is usually the enzyme of choice due to the presence of carrier RNA in most viral RNA preparations.

Procedure

With the optimized Sensiscript Reverse Transcriptase reaction buffer, tedious pipetting and pre-incubation steps are eliminated, and no additional RNase H digestion step is needed (see flowchart " Sensiscript Reverse Transcriptase Procedure").
See figures

Applications

Sensiscript Reverse Transcriptase is suitable for use in the following applications:

  • Standard RT and RT-PCR
  • Quantitative, real-time RT-PCR
  • Differential display RT-PCR
  • Primer extension and RACE analysis
  • Synthesis of double-stranded cDNA for cloning

Supporting data and figures

Specifications

FeaturesSpecifications
applicationscDNA synthesis, RT-PCR, qRT-PCR
reactiontypeReverse transcription
enzymeactivityReverse transcription
realtimeorendpointEndpoint
mastermixNo
sampletargettypeRNA template
singleormultiplexSingle
withwithouthotstartWithout hotstart

Resources

User-Developed Protocols (1)
The protocol has been used successfully for Cy3-, Cy5-, and biotin-labeling of cDNA from <50 ng of total RNA or poly A+ mRNA.
Kit Handbooks (1)
Sensiscript Reverse Transcriptase for First-strand cDNA synthesis using <50 ng RNA; Two-tube RT-PCR; One-tube RT-PCR
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Quick-Start Protocols (1)
Certificates of Analysis (1)