Isolation and characterization of microsatellites in oil palm (Elaeis guineensis Jacq.)
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Date
2007
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ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD
Abstract
This study represents the first Indian effort in isolation of microsatellites
from Oil palm (Elaeis guineensis Jacq.). Microsatellites were isolated by the
selective hybridization method in which DNA fragments containing repeat motifs
specific to repeat probe used in hybridization were captured. In the present study
two biotinylated oligonucleotides of the Di and Trinucleotide repeat types were
used as probes in individual hybridization reactions.
Genomic DNA was isolated from flushing tender leaves and digested with
RsaI restriction enzyme. To the digested DNA, Super SNX ds linkers were
ligated. The linker ligated genomic digest was hybridized against biotinylated
repeat oligonucleotide and eluted microsatellite enriched DNA fragments were
amplified by PCR using Super SNX primer. Enriched fragments were cloned in
pGEM- T cloning vector and transformed into DH-5α competent cells. Totally,
100 white colonies were screened and the clones having insert size above 300 bp
were selected and sequenced. Of the 30 clones sequenced, 9 were found to be
positive for microsatellites. Out of 9 microsatellites 5 were dinucleotide repeats, 4
were trinucleotide and imperfect repeats. To the captured microsatellites, SSR
primers were designed and standardized.
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Keywords
ISOLATION, CHARACTERIZATION, MICROSATELLITES, OIL, PALM