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Genic-enriched Genomic Sequencing

   

In all higher plants that have been investigated, most of the repetitive DNAs are hypermethylated at 5′-CG-3′ and 5′-CNG-3′ residues. Considering this, genic enriched libraries were constructed from the six diverse genotypes of G. hirsutum by methylation sensitive restriction using ClaI and HpaII enzymes individually. These genic enriched libraries were sequenced using 454 GS FLX sequencer (Roche) and further explored for the genomic resources.

 
       
    Complete work plan including sequencing and bioinformatic analysis  
     
       
     
    Details of genotypes used for Genic-enriched library preparation        
   
Genotypes    Species     Source of collection Accession No.     Origin
JKC 703 G. hirsutum JK Agri., Hyderabad, Andhra Pradesh 250304 New World
JKC 725 G. hirsutum JK Agri., Hyderabad, Andhra Pradesh 250305 New World
JKC 737 G. hirsutum JK Agri., Hyderabad, Andhra Pradesh 250342 New World
JKC 770 G. hirsutum JK Agri., Hyderabad, Andhra Pradesh 250334 New World
LRA 5166 G. hirsutum JK Agri., Hyderabad, Andhra Pradesh 250341 New World
MCU5  G. hirsutum USA-D, Dharwad, Karnataka 250336 New World
     
             
    Selection of six diverse G. hirsutum genotypes based on AFLP genetic diversity     
 
             
    Restriction digestion of genomic DNA with methylation sensitive (BstBI, HpaII, and ClaI)
and in-sensitive (
HindIII and EcoRV) restriction enzymes