小金海棠总RNA提取方法比较及cDNA的LD-PCR扩增

用CTAB和SDS-酚两种方法提取了小金海棠根的总RNA,以CTAB法提取的总RNA为模板,用长距离PCR(LD-PCR)的方法合成了双链cDNA并进行了扩增。结果表明,CTAB法比SDS-酚法提取的总RNA纯度高、完整性好,宜于进行RT-PCR和cDNA文库的构建,SDS-酚法操作简单易行,提取产物可以直接用于转膜进行Northern杂交。

Comparison of Methods for Total RNA Isolation from Malus Xiaojinensis and cDNA LD-PCR Amplification

The total RNAs from roots of Malus xiaojinensis were isolated by the methods of CTAB and SDS-Phenol. Using the CTAB method, double-strand (ds) cDNA was synthesized and amplified by Long-Distance(LD) PCR. The results showed that CTAB method is more suitable than SDS-Phenol for isolating total RNA of Malus xiaojinensis, the CTAB method extracted more integrative and higher purity RNA for RT-PCR and cDNA library construction ,the procedure of SDS-Phenol method is more simple for isolating the RNA used to transferring to nylon membrane for Northern blot.