甘蓝型油菜银染mRNA差别显示条件的建立及优化

Establishment and Optimization of Sliver Staining Differential Display of Brassica napus L.

mRNA差异显示技术是分析差异表达基因的强有力的工具。本实验针对油菜差别显示体系筛选优化,最终获得了差别显示体系。研究结果为:RNA模板在2μg,反转录产物用量为1.5μl时,可以扩增出较多且清晰的条带;PCR退火温度在40℃扩增条带清晰。该体系用于甘蓝型油菜低磷胁迫前后基因差异表达研究稳定可靠。

mRNA differential display is a powerful instrument to indentify differentially expressed genes. This experiment is designed to optimize a differential display with silver staining of oilseed, and a optimized differential display system is obtained. The results showed ：Many and clear bands were observed ,When the amount of RNA used was about 2μg and the amount of reverse transcription products used was 1.5μl. The annealing temperature on DDRT-PCR Amplification was 40℃ ,which produced many cDNA band. This system which was used for the analysis of gene expression under low phosphorus condition in Brassica napus L. is stable and reliable.