DNA提取
普罗索皮斯
基因组DNA
聚合酶链反应
色谱法
萃取(化学)
DNA
核酸
化学
乙醇沉淀
吸光度
液氮
浸渍(污水)
生物
植物
生物化学
基因
材料科学
复合材料
有机化学
作者
Claudia Yared Michel-López,Daniel González-Mendoza,Onécimo Grimaldo-Juárez
标识
DOI:10.4238/2013.september.27.10
摘要
The extraction of high-quality genomic DNA from Prosopis spp for polymerase chain reaction (PCR) amplification is complicated, owing to the presence of a high percentage of secondary metabolites that bind to or co-precipitate with nucleic acids.In the present study, we report a modified sodium dodecyl sulfate/phenol protocol that eliminates the use of liquid nitrogen in the maceration process, β-mercaptoethanol in the buffer extraction, and the ethanol precipitation step.The A 260 /A 280 absorbance ratios of the isolated DNA were approximately 2.0 to 1.9, suggesting that the DNA fraction was pure and can be used for further PCR analysis.The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction.Finally, this proposal represents an alternative fast, cheap, and effective method for the isolation of genomic DNA from fresh leaves of Prosopis spp, even in low-technology laboratories.
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