Can anyone explain the step up PCR? The following is the paragraph, I could not understand the PCR condition for step up PCR
[For the DNA-dependent RNA polymerase II second largest subunit (RPB2), primer-pair RPB2-5F and RPB2-7Cr (Liu et al. 1999) was used with a step-up PCR that started with 5 cycles and annealing temperature of 48 C, followed by 5 cycles at 50 C and a final 25 cycles at 52 C. ]
Thanks
The polymerase chain reaction (PCR) is a powerful method for fast in vitro enzymatic amplifications of specific DNA sequences. PCR amplifications can be grouped into three different categories: standard PCR, long PCR, and multiplex PCR. Standard PCR involves amplification of a single DNA sequence that is less than 5 kb in length and is useful for a variety of applications, such as cycle sequencing, cloning, mutation detection, etc. Long PCR is used for the amplification of a single sequence that is longer than 5 kb and up to 40 kb in length. Its applications include long-range sequencing; amplification of complete genes; PCR-based detection and diagnosis of medically important large-gene insertions or deletions; molecular cloning; and assembly and production of larger recombinant constructions for PCR-based mutagenesis . The third category, multiplex PCR, is used for the amplification of multiple sequences
that are less than 5 kb in length. Its applications include forensic studies; pathogen identification; linkage analysis; template quantitation; genetic disease diagnosis; and population genetics.
Dear Ajonok Hano,
I think
If you know how to set up TD-PCR , you can do the same thing at TU-PCR
Here is a protocol for you :
Upasana Kachroo
Owayes M. Hamed Leila Hosseinzadeh Anvar What would be the protocol in my case? In standard PCR, we are going like this,,, 95, 94, 50-60, 72, In this case how will I adjust PCR protocol? Thanks
Reference is made to the conventional PCR where the gradient of the whole block shifts after different cycles.
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