cDNA libraries - (Nov/27/2006 )
hello every one
i prepared cDNA libraries from HEK cell and i want to clone it in to vector i need to know is there any strategy for this or any one how do it to help me
You've already got your cDNA library constructed and cloned in E. coli, ya? Or do you only have cDNAs (or mRNAs) and wish to clone them but unsure of which vector system to choose?
If it's the latter, you can try out looking for "cDNA library construction kits" in google. It'll normally direct you to Biocompare and you'll be able to search for the system you wish to try.
If you're cloning only a single (gene specific) cDNA, e.g. actin, well... you can try T/A cloning or RE cloning (or blunt end cloning). The vector should depend on what you want to do with your cloned cDNA later. If it's for cloning (or sequencing) sake, any cloning vector should be alright. If you're going to do expression (in prokaryotic or eukaryotic system), you should be looking for expression vector.
It's best to try a system (or manufacturers) that your lab has in succeeded using it previously.
i did what you say with RE cloning but when i transformation i did'n have any colony and i'm sure my competent cell fine but i don't know why . and by the way i whant to know the perfection size of cDNA library
thank you very much