Please tell me how to add BSaI site in my grna oligo. As vector has BsaI site. when i cut with enzyme for the ligation it should be in my oligos also.
As BSaI seq is this
5'GGTCTC3'
3'CCAGAG5'
if my oligo are 5'XXXXXXXXXX3'
3'XXXXXXXXXX5'
please tell me which nucloetide i have to add in my oligo to insert in vector.
Robert Adolf Brinzer
You need to add the restriction site at both ends of your oligo. Still directional cloning is advised as using only a single restriction enzyme does not guarantee your oligo will ligate in the correct direction.
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