Qiagen, 205211, Sensiscript RT Kit (50)

For 50 reverse-transcription reactions: Sensiscript Reverse Transcriptase, 150 µl 10x Buffer RT, 100 µl dNTP Mix (contains 5 mM each dNTP), 1.1 ml RNase-free water

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

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.

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.

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

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