用改良的TRIZOL法快速提取仿刺参和中间球海胆的总RNA

Rapid extraction of total RNA from sea cucumber Apostichopus japonica and sea urchin by the modified TRIZOL method

用改良的TRIZOL法和普通TRIZOL法分别提取仿刺参Apostichopus japonica体壁、肠、体腔液和中间球海胆Strongylocentrotus intermedius的管足、性腺、齿间肌的总RNA。结果表明:用普通TRIZOL法提取时间长,电泳条带不很清晰,存在降解且各个组织提取的状况不稳定,杂质量多;而用改良TRIZOL法能够快速提取仿刺参、海胆组织的总RNA,电泳后得到的28S rRNA和18S rRNA条带清晰、完整性好、纯度高、得率高,其A/A为2.04~2.14,总RNA浓度为44.96~115.02 ng/μL。

The total RNA was extracted from body walls,intestines,and colelomic liquid in sea cucumber Apostichopus japonicus with rich in collagen,polysaccharides and pigments and tube feet,gonad,muscles among teeth of the sea urchin Strongylocentrotus intermedius by improved TRIZOL one step method to replace the traditional TRIZOL method with long period,and unclear electrophoresis bands.The modified TRIZOL method was found to extract RNA in the sea cucumber and the sea urchin rapidly,and to have clear electrophoresis bands of 28S rRNA and 18S rRNA,good integrity,high purity,and high output ratio.The A260/A280 was varied from 2.04 to 2.14 and the condense between 44.96 and 115.02 ng/μL.The modified method is characterized by simply repeatable and steady operation,and less time.