Amplification of pig microsatellite markers using multiplex PCR

In order to rapidly amplify pig microsatellite markers and save materials,multiplex PCR was used and its reaction condition was optimized.Forty-six combinations of multiplex PCR with good effects were obtained.Thirty of them are duplex-PCRs,sixteen are triplex-PCRs.The results of multiplexes showed that the concentration of primers varied among 0.06～0.3μmol/L,the Mg~(2+) concentration among 1.5～3.0 mmol/L;0.2～0.4 U of Taq polymerase and 1.0-,1.2-,1.4-,1.6-fold buffer were used,the annealing temperature and the cycle number varied among 52～60℃ and 32～50℃,respectively.All multiplexes were further combined into 17 sets for the electrophoresis on ABI 377 sequencer.