I am confirming my single copy deletion in yeast with southern blotting.i have no issues with the technical part as my blots were fine but their might be problem with my probe or genomic dna or enzyme , I cant figure it out. I have calculated all the predicted bands in my wild type as well as transformed clones with different enzymes(ecor1,ecor5,pst1,)
, using differwnt probes(3'UTR and promoter) but some clones showed showed correct band size some shows an extra band of 3 kb which is also coming in my wild type. And this is the problem that wild typw also shows 1 n smtimes 2 extra bands which is coming in my transormed clones too.it should not be their at all.
What can be the problem I dont know?
Please help in suggesting somthing n how to solve this issue.
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