generating a cDNA library - (Apr/02/2008 )

Hello there, I´m planning to generate a cDNA library to make a yeast two-hybrid screening. I will be using the Matchmaker kit from Clontech, which, in order to generate the cDNA from your RNA, offers the possibility of using oligo-dT primers or random primers. Considering that the two-hybrid system is supposed to detect protein-protein interactions between the protein you use as a bait and the proteins that will be translated from your cDNA library, which option do you think would be the best one, using the random primers or the oligo-dT? Thank you very much for your help.

Making a cDNA can be very difficult and you may not get every expressed gene! Why not look for a commercial library which has gone through a quality control? If you can't afford one, ask around and see if any of the labs around you have a library. All you need is one ul and you can expand the whole thing pretty easily. I did a two hybrid (bacterial) from Stratagene and got the cDNA library already subcloned into the needed vector. Saved me a ton of time and trouble!!