Genome Sequencing Strategy

Here we have discussed a genome sequencing strategy which is commonly adapted in labs.

The steps in the complete genome sequencing will generate the cDNA sequences that can be used to develop BACs and cDNA microarrays. But since there limited genome data of the organism available, the sequencing will be carried out in two distinct stages.

 

Stage I:

In this stage, the cDNA library of the organism will be generated. The database of novel EST’s from the most diverse EST library would be prepared that can be used for gene annotation, cloning and development of microarray’s etc. The EST’s will expand the understanding of types of genes expressed in the organism . The genome size of the organism will be estimated by comparing the reassociation rate. The genome size will provide a significant information by which we aim to generate BAC genomic DNA library that will inform about GC content of the genome, repetitive sequences and level of polymorphism in the genome.

Stage II:

In this stage, whole genome random shotgun sequencing is proposed for the complete genome sequencing of the organism. This method utilizes comparative genomics protein coding and regulatory regions on the genome. The sequence-tagged BAC clones are mapped to the polytene chromosomes so as assembled scaffolds are directly localized in the genome of the organism  with specific orientations. We aim to produce a draft genome with the knowledge of genomic organization, to carry out the comparative genome analysis and the complete finishing of the genome is done. The finished genome is released to a public domain after getting approval by several committees.