Cloning two inserts at a time in a vector - Cloning two inserts at a time in a vector (Mar/23/2011 )
Hello every1... I am new in this cloning thing and I am working on my PhD project which which involves cloning. Can I insert my gene of interest + shRNA expression sequence within a same vector. The vector I am interested in is ZD55 (adenoviral based).
.. Waiting and thankx in advance...
Not 100% sure on your inserts but I know that Infusion cloning from clontech allows you to have a 15bp overhang on each of your inserts so you can insert multiple into one vector. I'd check out their kit, its pretty good.
I don't know about your specific vector, but usually you can express your gene of interest and shRNA from one vector. You may need two different promoters though, because mRNA is PolII and shRNA is PolIII promoter based.