Representative gel images on initial PCR screening of an ovine BAC library using comparative primers from the bovine sequences. Approximately 190,500 random BAC clones were organized into pooled super DNA plates of rows, columns, and plates to facilitate PCR screening. Location of a target positive BAC clone in the library was determined usually by two runs of PCRs, one for “plate” and the other for “row + column”. The procedure eliminated the need for hybridization-based screening with radioactive 32P labeling. Gel images of PCR screen band on (A): Row pool of P098 BAC plate using the primer pair S036; (B): Row pool of P056 BAC plate using the primer pair S109; (C): Row N of P083 BAC plate; (D): Row F of P162 BAC plate. M: DL2000. Sample: PCR Products. A ~ P: Number of Row. 1 ~ 16: Number of Column (only partial shown here). P: Positive control (The amplified PCR products using the sheep genome DNA as templates).
Li et al. BMC Genomics 2012 13:398 doi:10.1186/1471-2164-13-398