Comparison between Two PCR Techniques for Amplifying the 5′ Flanking Sequence of the rbcS Promoter from Glycine max

The research was conducted to compare two PCR techniques for isolating a target DNA segment flanking the known sequence.Inverse PCR(IPCR),with two pairs of specific primers,was performed using genome DNA as the template digested with EcoRI,followed by self-ligation.TAIL-PCR,with combination of three nested specific primers and an arbitrary degenerated primer,respectively,was performed using the same genome template.438 bp and 867 bp of specific fragments were obtained by IPCR and TAIL-PCR,Sequencing results confirmed primers and the known sequence.Comparatively,TAIL-PCR technique is very simple and reproducible and can obtain the target fragment in a shorter time.The results showed that TAIL-PCR is more efficient than IPCR in identifying the flanking sequence.