Problems by using pcDNA 3.1/Myc-His(+) B - (Jan/28/2008 )

Hello, everyone

Please give me some advice.

Recently, I have some problem by using pcDNA 3.1/Myc-His(+) B.

I am trying to cut Hind3 and Kpn1 cutting sites in order to insert my favorite gene.

Since I have tried several times and still can not get my favorite gene inserted, I was wondering

is it possible to cut pcDNA 3.1/Myc-His(+)B by Hind3 and Kpn1 at the same time?

Would the two restriction enzymes compete with each other since their cutting sits are near by?

Thank you very much~~

Check out the NEB catalogue or web site. They have tables on how many flanking bases are required for complete digestion with each restriction enzyme.

do you have the sequence of your plasmid?
If you can count the number of bp separating the two sites you can then determine if a double digest or even if the digest you can considering is possible.