I am currently [color=#0066FF !important]working[/color][color=#000000] on a project where we have received a BAC [/color][color=#000000]library[/color][color=#000000] with 100 clones pooled per well. My boss has said we will screen using PCR. We will do an initial screen with PCR, then for the wells which received a postitve reaction we will dilute then 1/96 and PCR again. We will then sequences using 454 the diluted [/color][color=#0066FF !important]positive[/color][color=#000000] wells. I cannot find any material online about this, and I am not sure if my supervisor knows an exact protocol. I am slightly confused about this whole method, is there anyone who can further explain it to me or provide me with a reference on this???[/color]
The only information I find is about superpooling and 3D plates etc. We are not using this, we only have the plates of 100 clones pooled per well.
[color=#000000]Any input is greatly appreciated, thank you.[/color]
Edited by Irrateelevatorguy, 08 February 2012 - 07:05 AM.