Abstract: Objective Fluorescent labeled universal primers PCR in two-steps was established to screen microsatellite loci of naked mole rat. Methods A fusion of the forward primer and universal primers in 5' end was created. The first PCR was conducted with reverse primers and forward primer extended with universal primers. The second PCR was initiated after the adding of fluorescent labed universal primers. With conventional fluorescent labeled primers PCR as a control, PCR product was detected by agarose gel electrophoresis and capillary electrophoresis. Results Fluorescent labeled universal primer PCR had strong specificity and bands in agarose gel electrophoresis was sharp. Furthermore, the product had high consistent polymorphism with those in traditional PCR, with size being about 15 bp. They were in line with expectations. After analysis of capillary electrophoresis, it also showel equal number of allele with these two different method. As well, this fluorescent labeled universal primer PCR had high amplification efficiency and being specific, operational. Conclusion The fluorescent labed universal primers PCR in two-steps was applicable to screen a high-flux of microsatellite loci in naked mole rat.

Key words: Naked mole rat, Screening microsatellite loci, Fluorescent labeled universal primers, PCR