Problems with Transformation / DH5-alpha / Plasmid DNA ??!!
Raghavendra V
Wednesday, 16 January 2008 16:10 UTC
We have cloned a gene, and transformed it into DH5-alpha E.coli cells with suitable antibiotic (of course in the right concentration). The problem is, after incubating O/N at 37 deg C, we notice very tiny colonies on the LB-agar plate, and with great difficulty if we are able to locate, and inoculate a single colony into LB for Miniprep, at the end of the procedure we do not find any plasmid DNA!! (on 1% TAE agarose gel!!) Can any of you give a solution for this problem?!
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hi aparna,
thanks for your feedback! i am using a chemically competent Stbl3 strain from Invitrogen. but why then my unpurified controls give many more colonies? this is what i do not understand… i am really eager to know how Raghavendra V could solve his problem with tiny colonies…thanks a lot,
mirzo
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