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Purification and Sequencing of Xb10 interacting library cDNAPrey
   The E. Coli culture after propagation was spun down to obtain the pallets. Lysis buffer (10% SDS, 0.2M NaOH) and NaOAC were added and the mixture spun at 14,000 rpms for 4 minutes. RNAse and PCI (50% phenol, 50% choloroform, 1% iso-amyl alcohol) were added and spun at 14,000 rpms for 8 minutes. Excretion of the supernatant was followed by the addition of ethanol. The sample was spun again at full speed and autoclaved water was added to the samples.
The cDNA fragments (suspended in water) obtained from purification were loaded into a gel box with agarose gel. The system was allowed to run for 1.5 hours with 40 lanes of cDNA and 2 markers. ( Sombrook et al, 1993)
     Xbi-2m-2, xbi-1c-1, xbi-2c-1, xbi-3c-2, xbi-4c-1, xbi-5c-1, xbi-6c-1, xbi-8c-1, xbi-9c-1, xbi-10c-1, xbi-11c-1, xbi-12c-1, xbi-14c-1, xbi-18c-1, xbi-19c-1, xbi-20c-1, xbi-22c-1 were selected for sequencing via the gel picture obtained. DNA lanes that appear to have different size fragments were sequenced (DNA purification kit used) (Chern et al., 2001)
cDNA segment
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