Guys I am amplifying a specific locus on a legume genome. I get very faint bands and these product need a further digestion in order to reveal the polymorphism that I need to genotype my material. I get nothing after digestion. I PCR, run the undigested product (i.e very faint bands) and then digest. Is there anything I can do to improve my bands intensity? The PCR is optimized at 94 for 1 min, 35 cycles of 94 1 min, 57 1min and 72 2min and finally and extention of 72 5min. My fragments are 1.0 kb and 1.5 kb (after digestion). I use 30 ng DNA and primers and Promega mastermix to a total of 25 ul.
can someone please suggest something. kind regards Lebo RSA _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
