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Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples

机译:改良盐分致密荒地土壤样品群落DNA提取方法的改进

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Graphical abstract Display Omitted Abstract 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 4 )SO 4 . 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 4 )SO 4 is crucial step in humic acid removal from extracted DNA samples for soil samples containing high salinity and clay particles.
机译:为了克服盐引起的干扰以及裂解所需的细菌细胞释放中的致密性的问题,Yates所描述的方法。 (1998),通过添加Al(NH 4)SO 4从土壤微生物群落中分离基因组物质(脱氧核糖核酸,DNA)进行了优化。在所测试的样品中观察到非常低的总存活数,因此主要需要使用大量土壤来从荒地土壤中分离DNA。如果由于土壤中存在致密的性质和盐分浓度,可商购的珠击和酶解方法无法分离出任何数量的社区基因组DNA,则该方法证明了自己的有效性。 ?该协议被发现对于用于土壤微生物群落DNA提取的高粘土含量土壤样品有效。 ?裂解温育和土壤量的变化可能有助于微生物含量低的土壤样品。 ? Al(NH 4)SO 4的添加对于从提取的DNA样品中去除腐殖酸是至关重要的一步,而DNA样品则含有高盐度和粘土颗粒。

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