[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 approximately 0.3 micromol/L, the Mg(2+) concentration among 1.5 approximately 3.0 mmol/L; 0.2 approximately 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 approximately 60 degrees and 32 approximately 50 degrees, respectively. All multiplexes were further combined into 17 sets for the electrophoresis on ABI 377 sequencer.
Guo
Xiao-Ling
XL
College of Animal Science, Zhejiang University, Hangzhou 310029, China. xguo@email.com
Xu
Ning-Ying
NY
Looft
Christian
C
Reinsch
Norbert
N
Kalm
Ernst
E
D000818:Animals; D017931:DNA Primers; D005838:Genotype; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013552:Swine / Q000235:genetics*; D013696:Temperature
D017931:DNA Primers