Zaveri Purvi, Patel Rushika, Patel Meghavi, Sarodia Devki, Munshi Nasreen S
Institute of Science, Nirma University, Sarkhej-Gandhinagar Highway, Ahmedabad 382 481, Gujarat, India.
MethodsX. 2017 Jan 17;4:63-67. doi: 10.1016/j.mex.2017.01.002. eCollection 2017.
To overcome the issue of interferences by salt and compactness in release of bacterial cell required for lysis, method described by Yeates et al. (1998), was optimized for isolation of genomic material (Deoxyribo Nucleic Acid, DNA) from soil microbial community by addition of Al(NH)SO. Very low total viable count was observed in the samples tested and hence use of higher amount of soil is required primarily for DNA isolation from wasteland soils. The method proves itself efficient where commercially available bead beating and enzymatic lysis methods could not give isolation of any amount of community genomic DNA due to compact nature and salt concentrations present in soil. •The protocol was found efficient for soil samples with high clay content for microbial community DNA extraction.•Variation in lysis incubation and amount of soil may help with soil samples containing low microbial population.•Addition of Al(NH)SO is crucial step in humic acid removal from extracted DNA samples for soil samples containing high salinity and clay particles.
为克服盐分干扰以及裂解所需细菌细胞释放过程中的致密性问题,对Yeates等人(1998年)描述的方法进行了优化,通过添加硫酸铝铵从土壤微生物群落中分离基因组物质(脱氧核糖核酸,DNA)。在所测试的样品中观察到极低的总活菌数,因此主要从荒地土壤中分离DNA需要使用更多量的土壤。在商业上可用的珠磨法和酶解法由于土壤的致密性质和盐分浓度而无法分离出任何数量的群落基因组DNA的情况下,该方法证明了自身的有效性。•该方案对于高粘土含量的土壤样品用于微生物群落DNA提取是有效的。•裂解孵育和土壤量的变化可能有助于处理微生物数量低的土壤样品。•对于含有高盐度和粘土颗粒的土壤样品,添加硫酸铝铵是从提取的DNA样品中去除腐殖酸的关键步骤。