Krasova-Wade T, Neyra M
Laboratoire Commun de Microbiologie Centre de Recherche de Bel Air, Dakar, Senegal.
Lett Appl Microbiol. 2007 Jul;45(1):95-9. doi: 10.1111/j.1472-765X.2007.02149.x.
The aim of this study was to optimize DNA extraction from legume nodules to obtain large amounts of high-quality genomic DNA.
Nodules of different legume species were used. Varied concentrations of guanidine thiocyanate (from 6 mol l(-1) to 0.05 mmol l(-1)), a component of DNAzol, were tested. The quality of DNA extract was determined by PCR-RFLP. The best results were obtained with 0.5 mmol l(-1) guanidine thiocyanate, which resulted in greater DNA yield than with higher and lower concentrations or with DNAzol.
The procedure using 0.5 mmol l(-1) guanidine thiocyanate yields the highest DNA amount when compared with previously described protocols and offers a reliable method to isolate DNA from nodules of different origins.
Irrespective of nodule origin, DNA yield was increased significantly, by two (e.g., Vigna nodules) to seven (Acacia auricoliformis nodules) times. In addition, the proposed procedure's costs are lower than those using the DNAzol.
本研究的目的是优化从豆科植物根瘤中提取DNA的方法，以获得大量高质量的基因组DNA。
使用了不同豆科植物的根瘤。对DNAzol的成分硫氰酸胍的不同浓度（从6 mol l(-1)到0.05 mmol l(-1)）进行了测试。通过PCR-RFLP测定DNA提取物的质量。使用0.5 mmol l(-1)硫氰酸胍获得了最佳结果，其产生的DNA产量高于更高和更低浓度或使用DNAzol时的产量。
与先前描述的方案相比，使用0.5 mmol l(-1)硫氰酸胍的方法产生的DNA量最高，并提供了一种从不同来源的根瘤中分离DNA的可靠方法。
无论根瘤来源如何，DNA产量显著提高，提高了两倍（如豇豆根瘤）至七倍（大叶相思根瘤）。此外，所提出方法的成本低于使用DNAzol的成本。
