I was making the solution of SS DNA. During this I have to sonicate to make working solution. This solution is not working for yeast two hybrid. I have changed all the yeast strain, DNA and medium but still I didnt get the result this time with same protocol (That I used earlier). So can any body suggest me to check the quality of SS DNA, or any other remedies for this problem.
Which system you are using for yeast two-hybrid...? I mean the bait and prey yeast strains, vectors...? for salmon sperm ss DNA, check the concentration of the DNA and use as mentioned in the protocol. before using the ss DNA, denature it at 95 degree C for % min and therefter keep at 4 degree. after transformation store the leftover ss DNA at -20 or-40.
In yeast transformation, ssDNA is very seldom (never in my 15+ years of YTH experience) the cause for a total failure. Although a better preparation is certainly needed to achieve the highest possible efficiency, I would only be concerned about it's quality if a control experiment done in parallel (same yeast prep, PEG/LiAc, plasmid and plates - but different ssDNA source) have pinpointed the problem exactly on ssDNA... Is it the case?
Thanks Mr. LLYa SEREBRIISKII: I have planned the same experiment to test this, but i want to know either there is any direct method (spectrophotometer, nano drop etc) to test SS DNA, I am using Sigma SS DNA from salmon sperm, herring testes. I will do the technique also that Mr. Spencer has suggested. Thanks Mr. Spencer
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