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The Lift Pool Method For Isolation Of Cdna Clones From Lambda Phage Libraries, Janine M. Leblanc-Straceski, Pablo Sobrado, Sharon Betz, Julie Zerfas, Karen Morgan

Biology Faculty Publications

A PCR based strategy was developed to screen a Xenopus oocyte λgt10 cDNA library. The PCR-based lift pool (LP) method follows the same two tiered strategy as conventional screening of phage libraries by filter hybridization. Two rounds of plating, one at high density to detect the clone, and one at low density to purify the clone to homogeneity, are performed. In the first round, lysates from high density plates, termed plate pools (PP), serve as template for PCR. In the second round, phage particles adhering to plaque lifts of low density plates are washed off nitrocellulose filters to create LPs, …