Project Number: 6402-31000-008-05
Project Type: Reimbursable

Start Date: Apr 14, 2005
End Date: May 31, 2007

Objective:
The objective of this cooperative research project is to identify DNA clones containing the catfish immunoglobin heavy chain locus, sequence these clones, and establish the gene structure in this locus.

Approach:
Clones from a bacterial artificial library containing 150-200,000 base pair inserts will be identified by PCR-amplification screening. The BAC clones will be fragmented and subcloned into a plasmid vector. The plasmid subclones will be arrayed in 96-well format and sequenced using high-throughput technology. Sequences will be assembled using computer algorithms. Sequence information from individual clones will be used to design PCR primers to identify subsequent clones containing flanking genomic DNA, and the process will be continued until the entire locus (approximately 700,000 base pairs) is sequenced.