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Objectives
- Validation of
desired QTL in population
- Amplification
of yield locus
- Comparative
studies on sequences
Progress
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Contig assembly:-
Overgo probes were designed for yld 1.1a region of the QTL yld1.1. The
BAC library was screened with these overgo probes and BAC clones that
were selected were then determined their BAC overgo relationship. All 69
positive BAC clones we assembled into a contig.
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Sequencing of BAC
clone:- From the clones that have been designated into contig, a BAC
shotgun library is being constructed to facilitate the sequencing of an
entire BAC clone. Fingerprinting work has to be continued to fill in
gaps that have been found between the two. The array of clones into
contigs will then be confirmed by IMAGE and FPC. The BAC clone 24B23,
(approx 180kb) was selected for full length sequencing purposes. The
cloned DNA was purified and went through enzymatic and transposon
insertion procedures to obtained approx 1700 sequence reads for 5X
coverage of the clone. This 1700 sequence reads were generated by
digesting the BAC clone with 4 different restriction enzymes separately
and later cloning them for purpose of sequencing. Transposons were added
also to enable sequencing of gaps through the use of transposon primers.
The sequences were then assembled into their contigs. Sequence assembly
yielded a total of 50 contigs. The finishing phase to fill gaps is
currently ongoing.
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Genome databases and bioinformatics tools being
used in this study
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