Bhau B S, Gogoi G, Baruah D, Ahmed R, Hazarika G, Ghosh S, Borah B, Gogoi B, Sarmah D K, Nath S C, Wann S B
Plant Genomics Laboratory, Medicinal Aromatic & Economic Plants (MAEP) Division, CSIR-Northeast Institute of Science & Technology (CSIR-NEIST), Jorhat 785006, Assam, India.
Plant Genomics Laboratory, Medicinal Aromatic & Economic Plants (MAEP) Division, CSIR-Northeast Institute of Science & Technology (CSIR-NEIST), Jorhat 785006, Assam, India.
Food Chem. 2015 Dec 1;188:264-70. doi: 10.1016/j.foodchem.2015.05.004. Epub 2015 May 2.
Different species of Cinnamomum are rich in polysaccharide's and secondary metabolites, which hinder the process of DNA extraction. High quality DNA is the pre-requisite for any molecular biology study. In this paper we report a modified method for high quality and quantity of DNA extraction from both lyophilized and non-lyophilized leaf samples. Protocol reported differs from the CTAB procedure by addition of higher concentration of salt and activated charcoal to remove the polysaccharides and polyphenols. Wide utility of the modified protocol was proved by DNA extraction from different woody species and 4 Cinnamomum species. Therefore, this protocol has also been validated in different species of plants containing high levels of polyphenols and polysaccharides. The extracted DNA showed perfect amplification when subjected to RAPD, restriction digestion and amplification with DNA barcoding primers. The DNA extraction protocol is reproducible and can be applied for any plant molecular biology study.
不同种类的樟属植物富含多糖和次生代谢产物,这会阻碍DNA提取过程。高质量的DNA是任何分子生物学研究的先决条件。在本文中,我们报告了一种从冻干和未冻干叶片样本中提取高质量和高产量DNA的改良方法。所报告的方案与CTAB法不同,通过添加更高浓度的盐和活性炭来去除多糖和多酚。从不同木本植物和4种樟属植物中提取DNA证明了改良方案的广泛实用性。因此,该方案也在含有高水平多酚和多糖的不同植物物种中得到了验证。提取的DNA在进行RAPD、限制性酶切和DNA条形码引物扩增时显示出完美的扩增效果。该DNA提取方案具有可重复性,可应用于任何植物分子生物学研究。