摘要
采用多重PCR的方法,以快速扩增微卫星标记和节约试剂为目标,对其反应条件进行优化后,获得了46个扩增效果理想的微卫星标记多重PCR组合,其中30个为二重PCR,16个为三重PCR。实验结果表明,这些多重PCP反应的引物浓度为0.06～0.3μmol/L,Mg2+的浓度变化范围为1.5～3.0mmol/L,采用的PCR缓冲液的倍数为1.0、1.2、1.4或1.6,每个PCR反应聚合酶的用量在0.2和0.4U之间,退火温度及反应循环数分别为52～60℃和32～50℃。所有多重PCR进一步合并为17个可在ABI337测序仪上进行电泳的组合。
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.
出处
《遗传》
CAS
CSCD
北大核心
2004年第1期40-44,共5页
Hereditas（Beijing)
基金
德国H.WILHELMSCHAUMANN基金会资助~~
