I have transformed my DNA using pGEMT vector in E. coli. I have Nde1 and Xho1 as my up and downstream overhangs. My question is: Do i have to do digestion using EcoR1 alone first, streak another plate and send that for sequencing to confirm my insert? Then, i can do double digest with Nde1 and Xho1?
I went ahead and did double digestion without going the EcoR1 route. I have attached the results of the gel for that, were lane one is 1kb ladder, lane to is control without the enzymes, and the rest are my samples.
Can anyone help please???????