I am screening the clones obtained after successfully transecting the targeting vector into ES cells. After neomycin selection, the cells were expanded and Now, I am planing for screening to detect positive clones. Does any alternative molecular methods except southern blot and PCR, which helps in screening positive clones?
You could try designing a probe and doing DNA FISH? That would let you see if you have multiple insertions too.
Extract the vector (it is not inserted into the chromosome), make a linear form with restriction enzyme and make sure it is your interested size by agarose gel analysis.
You could try designing a probe and doing DNA FISH? That would let you see if you have multiple insertions too.
@Jorg: From the question I infer that there are at least two genes (GOI + Neo) that have been inserted into the genome. Oligos + labelling kit and away it goes. Now, did you want to help answer Praveen's question or did you just want to aim a heavy dose of sarcasm at me?
From a practical point of view there are no alternative methods to screen positive clones. You should go to screen by PCR and then confirm by southern blot.
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