Qiagen, 205111, Omniscript RT Kit (50)

For 50 reverse-transcription reactions: 200 units Omniscript Reverse Transcriptase, 150 µl 10x Buffer RT, 100 µl dNTP Mix (contains 5 mM each dNTP), 1.1 ml RNase-free water

Features

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

Performance

The high affinity of Omniscript Reverse Transcriptase (RT) results in highly specific and sensitive RT-PCR, even with low-copy transcripts (see figure " Sensitive RT-PCR of ≥10 copies "), and the ability to read through even complex RNA secondary structures without adjusting temperature or reaction conditions (see figure " Comparison of various reverse transcriptases ").

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 Omniscript RT Kit makes reverse transcription virtually trouble-free.

Principle

Omniscript Reverse Transcriptase (RT) has a high affinity for RNA, which enables efficient and sensitive reverse transcription of any template, leading to high yields of cDNA. It is provided ready-to-use with dNTPs and in an optimized reaction buffer that, together with the high affinity of Omniscript RT for RNA, enables read-through of templates with high GC content or complex secondary structure. Please note that the primer mix is not provided.

Omniscript RT is specially designed for all reverse transcription with any amount of RNA from 50 ng to 2 µg per reaction. Omniscript RT is also usually the enzyme of choice with viral RNA due to the presence of carrier RNA in most viral RNA preparations. In comparative experiments, Omniscript RT consistently outperforms other reverse transcriptases over a wide range of starting RNA amounts.

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

Applications

- cDNA synthesis for end-point PCR
- Synthesis of double-stranded cDNA for cloning
- RACE (Rapid Amplification of cDNA Ends)
- Linear RNA amplification
- Exponential RNA amplification
- SAGE (Serial Analysis of Gene Expression)
- Labeling for microarrays
- Analysis of transcription start site by primer extension

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