Gunnaporn - Veerachato
vgunnapo at MAILCITY.COM
Sat Feb 20 23:37:27 EST 1999
I used to have the problem of the 2 adjacent restriction sites when I used pET16b. The
restriction sites are NdeI, XhoI and BamHI, respectively and I wanted to insert my PCR fragment at
NdeI and BamHI sites. I could not ligate my PCR fragment into the vector. I solved the problem by
inserting another PCR fragment that has BamHI at both ends to extend to length between NdeI and BamHI restriction sites in my vector. Then I cut the vector with NdeI and BamHI. By this method I could make sure that the vector was actually cut and I successfully ligated my PCR fragment to it.
I hope this can help you to solve your problem.
vgunnapo at mailcity.com
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