3种不同方法对标本基因组DNA固定和抽提效果研究

Studies of the Influential Factors of Extraction of DNA from Fixed Samples Preserved with Three Different Kinds of Methods

利用常规基因组DNA抽提方法,对分别用甲醛、乙醇、聚乙二醇3种固定液固定的鲫(Carassius auratus)肌肉样品基因组DNA提取进行了比较研究。结果表明:5%甲醛保存的样品很难提取到完整的基因组DNA,乙醇95%和聚乙二醇(分子量为600)保存的样品均能成功提取出基因组DNA,并能成功地进行18S rDNA的扩增。同时探讨了不同固定方法对基因组DNA提取的影响。

Genomic DNA extraction from muscle samples of the crucian carp, （ Carassius auratus）, preserved with 5% formaldehyde, 95% alcohol and polyethylene glycol （PEG） （molecular weight： 600） are studied comparably in this paper. The results show that the genomic DNA can be extracted successfully from the samples preserved with 95% alcohol and polyethylene glycol （PEG） except the one with 5% formaldehyde by using commonly phenol/chloroform extracting genomic DNA method. The purified DNA, which are extracted from the samples fixed both with 95% alcohol and polyethylene glycol （PEG）, can be also applied to do amplifications by using universal prrmers （MX5 and MX3） in 18S rDNA, Moreover, the influential factors of extraction of DNA from samples preserved with different kinds of methods are discussed, and the quantity （ 173±4.48） of genomic DNA extracted from polyethylene glycol （PEG） and the one （160±3.5） from 95% alcohol are analyzed. Consequently, it can be inferred that polyethylene glycol （PEG） should be available as a good fixative for sample-preserving.