自制冰冻切片包埋剂在紫花地丁种子RNA提取中的应用

目的]通过比较使用自制包埋剂和合成胶水包埋紫花地丁种子提取RNA的效果,证明提取紫花地丁种子RNA时,使用自制包埋剂能获得高质量的RNA。方法]用自制包埋剂和合成胶水分别包埋紫花地丁种子,冰冻切片机切片破碎种子,提取RNA,通过RNA浓度、纯度及完整性的检测,比较2种方法提取RNA的效果。结果]自制包埋剂组和合成胶水组提取的RNA浓度分别为3.42和1.04μg/μl;1.0%琼脂糖凝胶电泳显示2组RNA都可见2条明显的条带,28S rRNA与18S rRNA条带亮度之比大于1,但合成胶水组有较为明显的拖带。结论]该试验结果显示,自制包埋剂包埋紫花地丁种子进行切片破碎,完全能满足提取RNA的需要,是一种成本低、效果好、操作简便、具有实用价值的方法。

Applying the Homemade Frozen Section Embedding Medium for Total RNA Extraction from the Seeds of Viola yedoensis

Objective] To extract the high-quality total RNA from the seeds of Viola yedoensis,the homemade embedding medium and synthetic glue liquid embedding medium were compared.Method] The seeds of Viola yedoensis were broken by frozen section method embedded by the homemade medium and the glue liquid.The concentration,purity and integrity of the total RNA were detected.Result] The RNA concentrations extracted by the homemade medium and the glue liquid medium were 3.42 and 1.04 μg/μl respectively.Agarose gel electrophoresis could see two distinct bands apparently.The brightness ratio of the 28S rRNA and the 18S rRNA bands was greater than 1.The total RNA extracted by the glue liquid medium method were smeared.Conclusion] The results showed that using the homemade frozen section embedding medium to break the seeds of Viola yedoensis method can meet the needs of most molecular biological experiments.It is a low cost,effective and simple method for high-quality RNA extraction.