The Chlamydomonas Bacterial Artificial Chromosome (BAC) library was prepared with genomic DNA from the cw92 mutant (strain CC-503). The DNA was partially digested with HindIII, then size fractionated and ligated into the HindIII site of a modified vector pBeloBAC11 (from Drs. H. Shizuya and M. Simon, Division of Biology, California Institute of Technology, Pasadena, CA (Kim et al.1996. Genomics 34:213-218)). The vector was modified by the insertion of genomic DNA containing the NAR2/NIT8 gene into the XhoI site of pBeloBAC11). The library consists of 40 dishes containing 384 clones each. DNA from all of the clones has been arrayed on nylon membranes, which can be obtained from GenomeSystems for hybridization with probes of interest. After positive clones have been identified, these can be obtained from Genome Systems (Incyte Genomics). The library has approximately eight fold coverage, so that on average (with more than 150 single copy probes tested) a probe representing a single copy gene will identify 8 BAC clones. The average size of the inserts in these clones is 75 kb (with a range of ca. 22 kb to 190 kb).

Techniques used for isolating BAC DNA, from Craig's Chlamy Corner