多寡核苷酸探针同步标记在Northern blot多基因分析中的应用

One-step labeling multi-oligonucleotide probes and detecting multi-gene simultaneously by Northern blot

目的:建立一种多探针同时标记同步杂交检测多基因的简化的Northern blot方法。方法:通过T4多聚核苷酸激酶5′末端放射性磷酸化标记法对Northern blot中多寡核苷酸探针同时标记,实现对同一张转移印迹膜上的多基因同步杂交显影检测。结果:与常规mRNA逐一检测的效果进行比较,同步简化法的各目的基因同时得到较好的检测。结论:同步简化法简化了实验过程,缩减了实验时间,节省了实验试剂,并使各基因表达丰度之间的比较具有更高的准确性,提高了Northern blot的检测效率和成功率。

Objective：To establish a simplified method for detecting muhi-gene simultaneously in Northern blot. Methods：Equal amounts of three oligonucleotide probes were mixed and labeled by 5＇termini phosphorylation with T4 polynucleotide kinase. Genes on the same membrane were hybridized and detected by three probes simultaneously. Results：Compared with routine single-probe detecting, our method was acceptable. Conclusion：The new method simplifies the experiment and saves time, whereas improves the comparing accuracy of each gene expression and enhances the effects of Northern blot.