摘要
用改良的CTAB法提取几个杏李品种成熟叶片的基因组DNA,裂解细胞核前先用洗液排除细胞质中的部分多糖、多酚、丹宁和果胶等物质的干扰;裂解细胞核时,在提取液中加入乙醇进一步去除多糖,结果表明,提取的基因组总DNA的纯度和完整性都较好,OD260/OD280值在1.70-1.93之间,DNA无降解现象,没有多糖污染,可被限制性内切酶消化,所提取的基因组DNA的SRAP扩增条带清晰,多态性好。说明用此方法提取到了质量合格的杏李基因组DNA,可以用于PCR和限制性内切酶分析。
The author have used the improved CTAB method to extract successfully the genomic DNA from mature leaves of some varieties of Prunus simonii. Firstly, before caryons lysis the washing buffer was used to avoid interference from amylose, polyphenol, tannin, pectin and so on in the cytoplasm; secondly, during caryons lysis, alcohol was mixed into the extraction buffer to further eliminate amylase. The results showed that the extracted DNA from the mature leaves by this method was pure and integral, the value of OD260nm/OD280nm was from 1.70 to 1.93, the degradation of extracted DNA was not found, the amylose in DNA was eliminated completely, and the DNA was suitable for digestion by restrictive endonucleases. SRAP analysis indicated the primer （ mel/eml ） produced clear polymorphic patterns. Therefore this method could be used to extract ideal DNA samples from the mature leaves of Prunus simonii Carr. and was suitable for PCR and restrictive enzyme analysis.
出处
《江西农业学报》
CAS
2008年第10期4-6,共3页
Acta Agriculturae Jiangxi
基金
河南科技学院重点科研项目(06029)
