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作 者:孙波[1] 鲍毅新[1] 张龙龙[1] 赵庆洋[1] 许婧[1]
出 处:《动物学杂志》2009年第6期145-150,共6页Chinese Journal of Zoology
基 金:浙江省自然科学基金项目(No.Y507080)
摘 要:利用微卫星引物在同一属、科、目不同种之间具有通用性的特点,通过PCR扩增、聚丙烯凝胶电泳和银染技术对社鼠(Niviventer confucianus)近缘物种大鼠(Rattus norvegicus)、小鼠(Mus musculus)中已知的70个微卫星位点引物进行跨种扩增,筛选适合社鼠相关研究的多态微卫星引物。结果发现,40个位点引物出现扩增条带,21个位点引物能够稳定扩增,其中15个位点杂合,13个位点具有多态性;PCR扩增的Mg2+浓度主要集中在1.5及2.0mmol/L,退火温度在50~60℃之间不等。虽然部分扩增产物有影子带的存在,但并不影响等位基因的判读。总体来看,利用大、小鼠的微卫星引物扩增社鼠的微卫星位点是可行的。Taking consideration of the fact that microsatellite primers can be amplified among different species in the same genus ,family and order,we used 70 Rat( Rattus norvegicus ) and Mouse( Mus musculus ) microsatellite primers to amplify their counterparts in a close species-the Chinese White-Bellied Rat( Niviventer confucianus ) through PCR amplification, polyacrylamide gel electrophoresis and silver staining technique. A total of 40 loci could be amplified in agarose gel electrophoresis, but after polyacrylamide gel electrophoresis, only 21 loci could be stably amplified, among which 15 loci showed hybridism and 13 loci showedpolymorphism. Mg^2+ concentration in PCR amplification was at 1.5 and 2.0 mmol/L, and the annealing temperature ranged from 50℃ to 60℃. The existence of stutter bands in partial amplified products did not affect the selection of purpose bands. Overall, it is feasible to amplify the Chinese white-bellied microsatellite primers by using their rat and mouse counterparts.
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