New answers tagged pcr
2
RNAses are enzymes, and there are various ways to inactivate them. Unfortunately, RNAses are rather stable proteins and autoclaving doesn't completely kill their activity. The common methods to inactivate are unspecific methods that will destroy any enzymes, either through covalent modification or degradation.
The most common methods for RNAse inactivation ...
1
You have to design your primers properly. Usually, in real-time PCR, you don't choose a very long product. Ideal product size is 150-300.
Next, see what your NRTI is analogous to. For e.g. if I am using AZT (Azathymidie), I would place my reverse primer at or after the last T.
There are alternate techniques as well. You can use primer extension ...
2
Reverse transcriptase (and most other commercial enzymes these days) is made recombinantly, then purified to varying degrees, using varying methods, among different suppliers (and sometimes between different lots from the same supplier). Some enzymes actually have multiple functional units combined in a complex, others may require a post-translation ...
1
What kind of reverse transcriptase is used in qRT-pcr is there a standard like taq dna pol is used for qPCR
M-MuLV (Monoley murine leukemia virus) reverse transcriptase
Some RT enzymes are engineered to remove the RNAseH domain so that template RNA persists for more rounds of RT.
Is it possible to use a biologically active Telemorease Elongation ...
Top 50 recent answers are included
