Wash at desired stingency. Placing the filters with care ie. Include fluorescent markers. You should get one to two positives per plate when probing a genomic library for single copy gene, cDNA libraries depend on the abundance of the mRNA you are screening for. Number each of the positives and set up eppendorf tubes with 1.0ml of TM10.
Accuracy lining up the key marks is crucial to get all the positives, whilst picking a small area.
Can gently vortex to increase phage dispersion. Infect 150 ul cells with 1 ul of 1 in 200 dilution of phage in TM, plate on small plate. Third round 1. Label nitrocellulose and carefully place into agar. Mark orientation by piercing through nitrocellulose and agar with needle and ink.