Development of Microsatellite Markers in Mango (Mangifera indica L.) using 5 anchored PCR

A 5' anchored PCR was utilized to develop microsatellite markers in mango. Di-repeat primers containing 3 or 7 degenerate bases at the 5' end of the primers were used to amplify genomic DNA flanked by simple sequence repeats. The PCR products were cloned to generate enriched microsatellite libraries. Clones containing microsatellite fragments were randomly selected and subsequently sequenced. A specific primer was designed and used in combination with a 5' anchored primer to produce a locus-specific microsatellite marker. The results indicated that 7 degenerated bases of 5' anchored primers reduced the efficiency of 5' anchored PCR. Sixteen microsatellite markers were developed and were able to reveal 46 alleles in 8 mango varieties.