青山羊皮肤组织差显快速银染技术的优化研究

STUDY ON OPTIMIZING THE FAST SILVER STAINING OF DDRT-PCR

差异显示PCR(differential display PCR,DDRT-PCR)技术是快速分离差异表达基因的强有力工具,但是其繁琐的染色程序和较高的假阳性率给研究者带来了不便。为了提高DDRT-PCR技术的效率和试验结果的清晰度和重复率,本研究以济宁青山羊皮肤组织的总RNA为试验材料,对影响DDRT-PCR的因素,如:反转录条件、反应体系和染色条件进行了优化研究。通过对染色程序和温度等染色条件的反复对比试验,建立了快速银染技术,得到了十分清晰的DDRT-PCR电泳图谱。快速银染技术既缩短了染色时间,又可以得到清晰的电泳图谱。

The method, of mRNA Differential Display PCR （ DDRT expressed genes. In order to improve the efficiency and repeatab -PCR） is a power tool to identify differential ility of the method, the skin tissues of Jining gray goat were used in this experiment, large numbers of contrasting and optimization test were carried on many factors which affecting the stability of DDRT - PCR, such as ： reverse - transcription condition, reaction system and dyeing condition. The technique of Fast Siver staining was established in this study. Finally, wer obtained an efficiency suitable method of RNA separated and more simple reaction system of DDRT - PCR.