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三种匀浆方式对线纹海马育儿袋总RNA提取质量的影响
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Abstract:
质量合格的RNA是RNA相关分子生物学实验的基础保障。线纹海马的育儿袋具有大量结缔组织，RNA提取过程中组织较难裂解。本研究探究了玻璃匀浆器研磨、磁珠震荡研磨和研钵液氮研磨三种不同的匀浆方式对总RNA提取质量的影响，结果显示，研钵液氮研磨法获得的RNA完整性、纯度及浓度较高，而玻璃匀浆器研磨、磁珠震荡研磨所提取的RNA浓度低、质量差。通过反转录PCR检测，通过研钵液氮研磨法得到的模版中可扩增出明亮、单一的目的基因条带。该研究为海马育儿袋总RNA的提取方法提供了参考。
High-quality RNA is the foundation of molecular biology experiments related to RNA. The brood pouch of Hippocampus erectus contains a large amount of connective tissue, which is difficult to be cleaved during RNA extraction. In this study, the effects of three different homogenizing methods, namely grind with glass homogenizer, beads shock and liquid nitrogen, on the quality of total RNA extraction were investigated. The results showed that RNA acquired by liquid nitrogen grinding method provided higher integrity, quality and concentration than that acquired by glass homogenizer and beads shocking. RT-PCR showed a single bright band of the target gene in the template acquired by liquid nitrogen grinding method. This study provides a reference for the total RNA extraction of brood pouch in Hippocampus erectus.
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