Construction of Whole Genome BAC map BAC by BAC Approach, RFLPs, STSs, SSRs, Whole-Genome Physical Maps

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Methods and Uses of Genomics and Proteomics Research Methods and Uses of Genomics and Proteomics Research - Introduction Genomics Research Methods for Whole Genome Sequencing Whole Genome Shotgun-WGS Sequencing Map First Sequence Later or Clone by Clone Strategy Construction of Whole Genome BAC Map / BAC-By-BAC Approach Building of Clone Contigs / Overlapping Series of Cloned DNA Fragments Whole Genome Sequence Data Electronic PCR (e-PCR) / Bridging the Gaps Between Mapping and Sequencing of Genomes From Genome Sequences to Function - Annotation

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Construction of whole-genome BAC map: BAC-by-BAC approach.

Maps having landmarks based on molecular markers like RFLPs, STSs, SSRs, and AFLPs are now available in all model animal and plant systems. These maps can be used for construction of whole-genome physical maps of ordered BACs. Following steps are involved in this exercise: (i) Fingerprinting of BAC clones having 10-15 times coverage of the genome is undertake. For instance, 300,000 BACs (each BAC = 150kb; 300,000 BACs = 45 billion base pairs) will give 15-fold coverage of human genome (3 billion 15-fold coverage of human genome (3 billion base pairs). This will give information regarding the number and sizes of fragment obtained from each BAC due to a specific enzyme.

(ii) On the basis of fingerprints and by locating the overlaps (restriction maps of BACs are also used, wherever required), BACs are assembled in contigs and singletons with the help of the software, FPc; physical mapping of contigs on the chromosomes is done using chromosome walking, fluorescence in situ hybridization (FISH), deletion stocks or radiation hybrids (for details on physical maps, consult) (iii) Contigs are joined wherever possible by examining their extreme ends and by gap filling approach discussed earlier in this; this leads to increase in contig size and the number o clones in an individual contig;

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