[Three steps for amplifying quick evolving region of DNA.].
A common problem in research of molecular evolution is difficult to efficiently amplify quick evolving target sequence of genes in different species or genus using specific primers, thus making experimental process and final analysis of total results delayed. Although using nested or semi-nested PCR can prominently increase PCR specificity, it really cannot efficiently amplify quick evolving region of DNA in our research of gene Fak56D. In this research, we need PCR products corresponding to gene Fak56D of different species of Drosophila melanogaster or other genus of Drosophila . For the high evolutionary rate,most materials did not produce qualified PCR products. To solve this problem, we initially used a combination method of three steps, i.e. semi-nested PCR taken together with orientational gel extraction, which satisfactorily met the demands of next cloning and sequencing steps.
Qiu
Gang
G
Faculty of Life Science, Hubei University, Wuhan 430062, China, Email: qiugang1997@163.com.
Zeng
Qing-Dao
QD
Jin
Shan
S
Qian
Yuan-Huai
YH
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004247:DNA