雀形目38种鸟类线粒体cytb的引物设计及PCR优化

Primers Design and PCR Optimization for Amplifying Mitochondrial Cytochrome b Gene of 38 Passerines

通过设计雀形目鸟类线粒体细胞色素b的通用引物并对其PCR反应体系进行优化,确定了雀形目38种鸟由基因组DNA扩增线粒体细胞色素b基因全序列的最佳反应条件.在10μl最佳反应体系中,dNTP浓度为100μM,引物浓度100nM,退火温度范围为51～55℃,模板量在50～150 ng之间,扩增反应效果最好.在上述最佳反应体系和条件下获得的目的条带清晰、明亮,可应用于雀形目鸟类分子系统学研究.

Through the design of primers for amplifying mitochondrial cytochrome b gene of 38 passerines and the optimization of PCR reaction volumes and conditions, the best conditions were got for amplifying whole sequences of mitochondrial cytochrome b genes of 38 passerines. Conditions including d NTP concentrations, primers concentrations and annealing temperatures were Optimized in this research. The results showed that in the best 10μl PCR volumes, d NTP concentration was 100μM, primers concentration 100nM, annealing temperature 51-55℃ and templates concentrations 50- 150 ng. Under the conditions above, the most bright and clear target bands were got after the agarose gel electrophoresis of PCR products. These reaction volumes and conditions can be used in molecular phylogenetic researches of passerines.