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CV2 is a subtracted library derived from a normalized eye library (nREE2), constructed according to the procedure described by Bonaldo, Lennon & Soares (Genome Research Genome 6: 791-806, 1996). For construction of the CV2 library, plasmid DNA from the normalized rat eye library was electroporated into competent bacteria for production of single-stranded circular DNA. This was then used as a tracer in a subtractive hybridization with a driver comprising: a) a set of about 1,000 arrayed clones from the non-normalized eye library CV0 (PCR amplified inserts from a plasmid DNA template preparation representing plates R-CV0-BRH through R-CV0-BRR). This represented 20% of the final driver population. b) A set of about 4,000 arrayed clones from the normalized eye library CV1 (PCR amplified inserts from a plasmid DNA template preparation representing plates R-CV1-BRS through R-CV1-BSC, R-CV1-BSE through R-CV1-BTC, and R-CV1-BV0 through R-CV1-BVU). This represented 80% of the final driver population.
Nucleotide
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