phosphorylation of primers using PNK - (Jun/22/2010 )
I phosphorylated my primers using T4 polynucleotide kinase - don't think it worked... Read somewhere that during the synthesis/desalting of primers they use ammonium hydroxide. Ammonium ions inhibit PNK. Any suggestions for removal of ammonium ions from primers?
1. The ammonium hydroxide is evaporated off during drying.
2. It takes a high concentration of ammonium to inhibit PNK. (Yes, I have performed the titration.)
You are unlikely to encounter the concentration levels necessary to inhibit the enzyme.