TAIL—PCR研究进展及其在畜牧兽医中的应用
设计引物、调整反应条件，增加操作的简便性；还可参考Suppression-PCR原理，选择引物和反应条件，获得较长的扩增片段。此外，TAIL-PCR可与多种技术联合使用，促使其在畜牧兽医中获得更广泛深入的应用。
关键词： TAIL-PCR；染色体步移；研究进展；应用
中图分类号： Q813 文献标志码：A 文章编号：2095-1191（2017）05-0926-07
Advances in TAIL-PCR and its application in animal
husbandry and veterinary
MA Ling 1， WANG Zhi-qiang 2，3， QIN Shao-min1， WU Jian-min1 *
（1 Guangxi Veterinary Research Institute / Guangxi Key Laboratory of Animal Epidemic Etiology and Diagnostic， Nanning 530001， China； 2 State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresearches， Nanning 530004， China； 3 College of Life Science and Technology， Guangxi University， Nanning 530004， China）
Abstract：Simple， rapid， high efficient and specific， thermal asymmetric interlaced PCR（TAIL-PCR）， a kind of chromosome walking based on PCR， was mainly used to isolate the flanking sequences of target DNA segments. The principle， advantage， disadvantage and reaction system optimization of TAIL-PCR as well as some critical problem were summarized. In animal husbandry and veterinary medicine， TAIL-PCR was mainly used to isolate exogenous gene insertion sites and clone flanking sequences of the known genes in transgenic animals. Being limited by the complexity of genome DNA， there were so many limited factors preventing cloning target genes from genome， such as amplification of nonspecific products caused by mismatch of specific primers and arbitrary primers， and the difficulties in obtaining positive products even using multiple degenerate primers caused by abnormal template GC content and highly repeated sequences. So the adjustments should be made depend on actual situation. When the templates were complex， adapter could be added to the primers to improve the specificity based on the principle of LM-PCR； while when the template were simple or the sequence were clear， primers could be designed and reaction conditions could be set based on the characters of the templates（high GC content） or known template sequence， which made it easier to operate； and the suitable primers and reaction conditions could be chosen to get longer amplification products based on the principle of Suppression-PCR. Moreover， TAIL-PCR could be used with other technologies to facilitate its application in animal husbandry and veterinary.