Short Note: Internal Size Standard for Microsatellite Genotyping
AbstractThe in-house production of fluorescently labelledinternal size standard offers the advantage of cost savingover the commercial size standard in microsatellitegenotyping. Based on the reported in-house internal sizestandard protocol, we have improved the method by generating21 DNA fragments (in a standard named as HM-400) with each size similar to that of the commercialsize standard. The consistent amplification of the correctfragment size was optimised via primer modulationfor non-templated nucleotide addition by Taq DNA polymerase.A total of six microsatellite loci were used toassess the accuracy of HM-400 and the mean standarddeviation of the size data was 0.19. The differencesbetween the fragment size means for samples sizedusing HM-400 and commercial size standard were smallwith an average of 0.29 bp. The production cost of HM-400 was only 10% of the cost of commercial size standard.