不同固定剂处理的对虾基因组DNA提取方法研究

Study on the DNA extraction from shrimp specimens by different methods

为了从乙醇固定和福尔马林固定的对虾标本中获得高质量的基因组DNA,对标本的预处理方法以及DNA的提取方法进行了研究。结果表明,75%乙醇作为固定剂更适合用于提取对虾标本的基因组DNA。对乙醇固定的对虾标本采用PBS溶液和TE溶液进行预处理,获得的DNA得率较高,改进法、DTT法和CTAB法是较为理想的DNA提取方法。对于福尔马林固定的对虾标本来说,所选用的3种预处理方法均能为后期的DNA提取创造条件,而4种提取方法中的DTT法和CTAB法,则较为适宜提取福尔马林固定的对虾标本DNA,其中,GTE溶液预处理与DTT法相结合为最优组合方案。

To improve the quality of the extracted genomic DNA from shrimp specimens kept in ethanol and formalin,pretreatment methods of specimens and methods for DNA extraction were studied.The results suggested that 75% ethanol was better to extract genomic DNA of shrimp specimens.The PBS solution and TE solution pretreatment methods contributed to improve DNA extration rate from specimens kept in 75% ethanol.The modified method,DTT method and CTAB method were more effective DNA extration methods.The three methods which were used to treat specimens kept in formalin helped to extract genomic DNA.DTT method and CTAB method were more effective methods for extracting genomic DNA from shrimp specimens kept in formalin.Combining GTE solution pretreatment with DTT method was the best combination method.