总RNA和mRNA来源的探针与cDNA芯片杂交的差异研究

The diverse research of the different probes when hybridize with the cDNA microarray which come from total RNA and mRNA

提取BEP2D细胞的总RNA并按两种方式进行cDNA芯片探针的标记,一种是将100μgBEP2D细胞的总RNA利用逆转录法直接标记成荧光探针,另一种是先从100μgBEP2D细胞的总RNA中分离出mRNA,然后再标记成荧光探针。将两份标记好的探针同时与含有230个基因的cDNA芯片杂交。杂交后的芯片经Axon4100B扫描仪扫描,发现两种方式标记探针的一致性为93.04%,并且mRNA来源探针杂交后的荧光信号值较总RNA的弱。探讨了这两种方法标记探针在基因芯片表达谱研究中的差异性,目的是为利用这两种方法标记探针进行基因表达谱研究者提供一些依据。

Total RNA from BEP2D cells was extracted and labeled by two different way,one was that100μg total RNA was labeled directly using RNA reverse transcript;another was that mRNA was isolated from100μg total RNA firstly and labeled using the same method.Then the two kind probes were hybridized with the cDNA microarray that containing230genes at the same time.After scaning using Axon4100B machine,the93.04%consistency was gotten between the two kind probes,and the probe fluorescent signal that come from mRNA is weak than that of come from total RNA.This study explored the difference of the two kind probes when hybridize with the cDNA chip,the aim is to provide some in-formation for the researchers who using this two kind probes.