[An optimized method for construction of genomic library].
Construction of genomic libraries is basic and important. Because of the laboriousness and high background of traditional methods for constructing genomic libraries, we improved them by overcoming these disadvantages. Two Ear I sites were chosen as the cloning sites, which can produce variable 3-base cohesive ends. Therefore the two overhangs could be devised to prevent a match and to avoid self-ligation of vector. Genomic DNA is cleaved partially with Sau3A I and subsequently incubated with dGTP and Klenow fragment of DNA polymerase Iso the self-ligation of fragments and ligation between them are blocked. In this study, the ARS probe vector (pHBM803/Trp) based on the improved method was constructed and then we constructed the Oryza sativa genomic library separately with the traditional method and improved method and compared them. The result of experiment indicated that the improved method could optimize the quality of library.
Xu
Song
S
Laboratory of Molecular Microbiology & Gene Engineering, College of Life Science, Hubei University, Wuhan 430062, China. xusong364@yahoo.com.cn
Zhang
Juan
J
Ma
Li-Xin
LX
D003001:Cloning, Molecular / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D018745:Genome, Plant; D015698:Genomic Library; D012275:Oryza / Q000235:genetics*