A total of 36 SSR loci from 30 CsLAC genes were selected for designing primers. To validate the primers, 45 tea cultivars or varieties were used for PCR amplification and subsequent resolution by electrophoresis. The reaction mixtures, thermocycling conditions and protocols for PCR product separation were performed based on a previous study [38 (link)]. The amplified fragments were separated on a 96-capillary automated DNA fragment analyzer (Fragment Analyzer™ 96, Advanced Analytical Technologies, Inc., Ames, IA). The separated DNA bands were visually scored using PROSize™ 2.0 software, which was included in the advanced Fragment Analyzer™ 96 system. Only one or two fragments were collected for each individual [37 (link)].
The number of alleles (Na), Shannon’s information index (I), observed heterozygosity (Ho), expected heterozygosity (He), genetic diversity (GD) and polymorphism information content (PIC) values were calculated with PowerMarker version 3.25 (http://statgen.ncsu.edu/powermarker/downloads.htm) (Liu and Muse 2005).
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