I want to clone ~100 different coding sequences from human or mouse into an expression vector to analyze phenotypes in tissue culture. I can do all of the steps, but I'm not sure what I should use as the template for my reactions. Ideally, I would purchase a pool of full length cDNAs from a bunch of organs, and amplify from that. Less ideally, I can generate cDNAs myself, from multiple organs and pool them. Does anyone have suggestions on a template source?
- Verify where these transcripts are generally present (this should not be difficult for common organisms)
- If they are all expressed in a certain tissue use cDNA from that tissue
- If not then pool the cDNAs from different tissues such that all genes can be amplified
- cDNA libraries (including whole organism) are also available and you may obtain them from the nearest repository (I guess NCBI also provides).
Some other things you can do:
Ask the primer synthesis company to provide primer sets it in a 96well format.
DNA synthesis is also a decent option (it takes something like $0.5 per base pair), but that can be expensive if your cDNAs are huge.