I want to order primer sets of RT-qPCR for miRNA detection based on the P K Busk et al paper ( without probe ) .
Is the DNA Reverse Phase Column purification method is proper for this aim ?
Laurence Stuart Dawkins-Hall
Dear Saeid
- In short yes they are more than suitable in grade
- However, for RT qPCR unless your primers are > 30bp ecen standard desalted purity are of sufficient quality: I routinely use all such primers at standard desalted grade
- For longer primers PAGE purified or a version of HPLC are used simply to ensure that that your full length primer is present @ > 90%. In this instance standard purity leads to a higher percentage of N-1 and N-2 truncated primers such that your full length might only approximate to ~ 70%
- But apart from elongated primers all levels of purity will work for RT qPCR
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