QIAGEN Omniscript and Sensiscript Reverse Transcriptases are unique recombinant enzymes with a high affinity for RNA, providing highly specific and sensitive reverse transcription without optimization.
| QIAGEN RT Kit Options | |
| Amount of starting RNA | Recommended RT Kit |
| 50 ng to 2 µg per reaction* | Omniscript RT Kit |
| <50 ng per reaction* | Sensiscript RT |
| * | The enzyme of choice depends on the total amount of RNA (including carrier) in the sample, regardless of the target RNA copy number. |
QIAGEN offers two recombinant reverse transcriptases with high affinity for RNA. This high affinity results in highly specific and sensitive RT-PCR even with low-copy transcripts, giving them the ability to read through even complex RNA secondary structures without adjusting temperature or reaction conditions (see figure "Comparison of Various Reverse Transcriptases").
Comparison of Various Reverse Transcriptases
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Reverse transcription was carried out with different reverse transcriptases according to suppliers＊ specifications, using the indicated amounts of total RNA from HeLa cells. 1/20 of the reverse-transcription reaction was used in a 25-cycle PCR amplification with QIAGEN Taq DNA Polymerase. A 1.7 kb beta-actin fragment was amplified. Omniscript: Omniscript Reverse Transcriptase (QIAGEN); MMLV RNase H每: RNase H每 reverse transcriptase from Moloney murine leukemia virus (Supplier L); MMLV: wild-type MMLV reverse transcriptase (Supplier L); AMV RNase H每: thermostable RNase H每 reverse transcriptase from avian myeloblastosis virus (Supplier L); AMV: wild-type AMV reverse transcriptase (Supplier P). |
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"). These difficult templates, however, prove no problem for QIAGEN reverse transcriptases (see figure "Full-Length RT-PCR Products"). With no need for optimization, Omniscript and Sensiscript RT Kits make reverse transcription trouble-free.
Lot-to-lot reproducibility of QIAGEN RT Kits is ensured by rigorous quality control at QIAGEN. The optimized Buffer RT, dNTPs, and water included in all Omniscript and Sensiscript RT Kits, are guaranteed RNase-free, and each lot of QIAGEN Reverse Transcriptases is thoroughly tested for RT-PCR reproducibility.
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RT-PCR was carried out with total RNA from maize leaves using the indicated RT reaction temperatures. A 1.2 kb fragment of the GC-rich maize gl2 transcript was amplified. A RT was carried out using the standard Omniscript RT protocol at the standard (37∼C) and higher reaction temperatures as indicated. B RT was carried out using the standard MMLV RNase H每 RT protocol (Supplier L) at the standard (42∼C) and other reaction temperatures as indicated. This standard protocol was carried out with the required preliminary denaturation step and the recommended RNase H digest step following RT. (Data from reference 1) |
QIAGEN Reverse Transcriptases are the ideal choice for gene-expression analysis and other applications such as:
Korfhage, C., Fisch, E., Schröder-Stumberger, I., and Oelm邦ller, U. (2000) Solving difficult template problems in RT-PCR. QIAGEN News 2000 No. 1, 12