基于PCR技术的人脑基因可变剪接差异性研究

Investigation of Difference of Alternative Splicing In Human Brain Based On PCR

目的:对人脑的基因可变剪接位点差异性进行分析。方法:通过RNA-Seq得到的转录组数据,分析推测出在不同年龄的7对样本的大脑前额叶皮层(prefrontal cortex,PFC)表达的某些基因具有可变剪接位点的差异性;为验证此推测,设计7对引物,提取不同年龄样本的RNA,通过反转录合成cDNA,采用PCR技术扩增目的序列。结果:在已剪接和(或)未剪接产物预期片段处有明显的条带。结论:通过Quantity One软件分析后,确定在不同年龄间存在可变剪接差异。基于PCR技术的基因可变剪接分析对RNA-seq数据集的预测结果进行验证是一种有效新颖的PCR新应用。

Objective： To investigate the aging related alternative splicing by PCR.Methods： Differences in splicing sites of the prefrontal cortex（PFC） in brain samples of 2 different ages were detected by transcriptome RNA-Seq data analysis.Seven pairs of primers were designed separately.The mRNA extracted from samples were mixed in equal amount,and then cDNA was synthesized by reverse transcription.Results： The target sequences were amplified.Conclusions： There were clear bands for different splicing pattern,established the existence of alternative splicing differences between ages.