2种PCR技术扩增微分离的蚕豆染色体

用接头介导的PCR (polymerase chain reaction, LA-PCR) 和简并寡核苷酸PCR(DOP-PCR)2种技术对微分离的蚕豆(vicia faba)大M染色体进行了扩增,并对扩增结果及2种PCR技术的优缺点进行了比较.结果表明,LA-PCR的扩增产物大小为0.3～3kb, 而DOP-PCR的扩增产物大小为0.3～1.3kb. LA-PCR的对照中未观察到扩增产物,而DOP-PCR的对照中却明显观察到扩增产物,此扩增产物是由于引物之间相互退火而形成的二聚体或更高级的聚合体. LA-PCR的Southern杂交信号明显强于DOP-PCR,可见LA-PCR的扩增效率明显高于DOP-PCR.

AMPLIFICATION OF MICRODISSECTED CHROMOSOMES FROM VICIA FABA BY TWO PCR TECHNIQUES

Linker adaptor mediated PCR (LA PCR) and degenerate oligonucleotide primed PCR (DOP PCR) are used to amplify the microdissected M chromosomes from Vicia faba . The results of amplifications and the advantages and limitations between the two techniques are compared. The results show that the PCR products of the LA PCR range in size from 0.3 to 3 kb while that of the DOP PCR range in size from 0.3 to 1.3 kb. In the control of LA PCR no amplification product is observed while in that of DOP PCR amplification product is observed obviously, which is dimer or higher polymer caused by annealing between primers. The Southern hybridization signal of LA PCR is stronger than that of DOP PCR, which indicates that the amplification efficiency of LA PCR is higher than that of DOP PCR.