HELP WITH RESTRICTION DIGEST OF PHAGE DNA - (May/16/2008 )
I NEED SOME HELP WITH RESTRICTION DIGEST OF PHAGE DNA. I HAVE CONCENTRATED MY PHAGES BY PEG AND NaCl METHOD AND PERFORMED DNA EXTRACTION BY PHENOL/CHLOROFORM METHOD. I GOT REALLY CLEAN BRIGHT BANDS ON MY GEL BUT AM HAVING TROUBLE WITH THE DIGESTS. I AM USING EcoR1, EcoRV, Dra!, Dde1, Hind111, Hinf1, BamH1 and Xba1, WITHOUT MUCH SUCCESS!!!
I HAVE ALSO USED THESE ENZYMES TO CUT LAMBDA DNA AND THERE IS NOR PROBLEM WITH THE ENZYMES THEMSELVES.
ANY HELP IN THIS REGARD WOULD GREATLY BE APPRECIATED!
ALSO, HAS ANYONE APPLIED PHAGES TO FOOD SAMPLES, AND IF SO, WHICH METHODS DID YOU USE...
I'd re-precipitate the DNA with ethanol. That should get rid of whatever is inhibiting your reactions.
Also, please don't write your messages in upper case. It looks like you're shouting at us... :-)
Same thought here. In fact, I find it difficult to read -wonder what may be the reason!
my sincere apologies for the upper case - its frustration!!!
No worries, we all know where you're coming from with that!