The Scriptum-55 reverse transcriptase is suitable for the synthesis of cDNA molecules from 100 bp to 12 kb in length. Stable at 55°C.
The scope of delivery includes the Scriptum-55 Reverse Transcriptase / Ribonuclease Inhibitor Mix as well as the "Ready to Use" 5x reaction buffer "complete". The buffer contains all necessary components like MgCl2 and dNTPs. You only need to add RNA and primers.
The reaction temperature of the reverse transcription is dependent on the structural properties of the RNA. The optimum temperature of the reverse transcriptase -55 is 50 ° C and is thus ideal for routine applications with simple and even difficult RNA templates.
An elevated temperature (maximum 55°C) should be used for highly structured RNAs. For very difficult templates we recommand the Bio&SELL reverse transcriptase 55 "improved".
In general, the optimal temperature and incubation time for each RNA should be adjusted. The easy and understandable protocol in our manual gives you important information and tips.
| Product | Article No. | Quantity |
|---|---|---|
| SCRIPTUM-55 Reverse Transcriptase | BS.56.500 | 50,000 units |
| BS.56.100 | 10,000 units |
Why the high operating temperature is so important:
The secondary structures of the RNA and its recovery after heating complicate the cDNA synthesis significantly. Initially, the RNA is heated for 10 minutes at max. 65 °C and thus the RNA linearizes and all loops are opened. During the "chill down" the reverse transcriptase is added to the reaction mix and the reaction starts. The reaction proceeds the better, the higher the operating temperature can be kept here. That is, because at a higher temperature the RNA formes no or less "loops". Because the reverse transcriptase has no "strand displacement", it will fall off the RNA in a "loop" and the sequences located behind the "loop" will be lost for a PCR product. These difficulties with the RNA and the loops in the cDNA synthesis can thus be avoided having a reverse transcriptase working at high temperatures, for example the SCRIPTUM-55 or the SCRIPTUM-55 improved.