Ligation problem

Gunnaporn - Veerachato vgunnapo at MAILCITY.COM
Sat Feb 20 23:37:27 EST 1999


Dear Andrew 

               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. 

Gunnaporn  Veerachato 
vgunnapo at mailcity.com  


Get your FREE Email at http://mailcity.lycos.com
Get your PERSONALIZED START PAGE at http://personal.lycos.com



More information about the Methods mailing list