ligation error

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ligation error

Post by nekonara » Sun Feb 17, 2013 2:48 pm

hello and good day !

i have done ligation of xylanase gene into pRSET B and pACYCDUET-1 vector

next , i incubate onto luria bertani (i use E. coli DH5a for host cell ) with antibiotic (amp50 for pRSET B, cam30 for pACYCDUET-1) at 37'C overnight.

on the next day, i found that only pRSET B have white colonies, but pACYCDUET-1 shows no colonies at all !

what is the possible reason? maybe you can answer it in point (short but easily to understand)

thanks for helping !

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Post by jonmoulton » Tue Feb 19, 2013 5:22 pm

It sounds like your transfection with pACYCDUET-1 did not work. Perhaps the cells were not competent.

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