Plant & Animal Genome IX Conference Plant & Animal Genome IX Conference
Poster: Large Insert Libraries, Gene Isolation, Etc.
P02_09.html
Sorghum is the world's fifth most widely grown cereal (Doggett, 1988; Martin et al, 1976). Sorghum is a diploid plant (2n=2X=20) with a haploid genome equivalent of 750 Mbp (Arumuganathan and Earle, 1991). In order to facilitate positional cloning, physical mapping and genome sequencing we have constructed a deep-coverage BAC library from Sorghum bicolor cv BTX 623. The library was constructed by partial digestion of genomic DNA with HindIII restriction enzyme and cloning into the HindIII site of the pCUGIBAC 1 vector. Random sampling of 576 clones indicated an average insert size of 115kb. The library is maintained in 288 384 well plates and was gridded onto 22.5X22.5 cm nylon membrane in duplicate by using a Genetix Q-BOT in a 4X4 array which resulted in 6 filters. Each set of 6 filters contains 110,562 clones in duplicate which represent 16X genome coverage. The deep coverage allows the isolation of a particular sequence with a probability of 99%. Screening of the filters with three different barley chloroplast probes and four rice mitochondrial probes indicates the library contains 0.69% chloroplast clones and 0.08% mitochondrial clones. For analyzing the synteny between the sorghum and monocot genomes the library was screened with 6 rice RFLP markers from chromosome 10 of rice (RGP) and 3 sugarcane markers link to rust resistance (CIRAD). Positive clones from hybridization have been identified and will be integrated into our sorghum fingerprint database.