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.