Nucleic acid protocols: Extraction and optimization.
Nucleic acid protocols: Extraction and optimization.
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DOI：
10.1016/j.btre.2016.10.001
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发表时间：
2016-12
期刊：
影响因子：
--
通讯作者：
Suo X
中科院分区：
文献类型：
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作者：
El-Ashram S;Al Nasr I;Suo X
一个简化的，半统一的，从不同的原核和真核来源提取DNA和RNA的协议。在裂解步骤期间，DNA和RNA处于三重保护（即EDTA、SDS和NaCl）下。在DNA和RNA提取后分别加入DNase和RNase。产量和质量是任何研究人员在核酸提取过程中的基本特征。在这里，我们描述了一个简化的，半统一的，有效的，和有毒的材料免费协议提取DNA和RNA从不同的原核和真核来源，利用核酸的物理和化学性质。此外，该方案表明，DNA和RNA在裂解步骤期间处于三重保护（即EDTA、SDS和NaCl）下，并且该环境不适合于RNA酶使DNA从RNA中释放，甚至不适合于DNA酶降解DNA。因此，当在DNA提取后加入RNA酶时，在RNA酶的影响下实现了RNA的完全去除，这对于任何方案都提供了最佳质量。类似地，分离的RNA中的DNA污染物被DNA酶降解以获得高质量的RNA。我们的方案在简单性、恢复时间、环境安全性、数量、纯度、PCR和RT-PCR适用性方面是首选方案。
A simplified, semi-unified, protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources. DNA and RNA are under triple protection (i.e. EDTA, SDS and NaCl) during lysis step. Adding DNase and RNase after DNA and RNA extraction respectively. Yield and quality are fundamental features for any researchers during nucleic acid extraction. Here, we describe a simplified, semi-unified, effective, and toxic material free protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources exploiting the physical and chemical properties of nucleic acids. Furthermore, this protocol showed that DNA and RNA are under triple protection (i.e. EDTA, SDS and NaCl) during lysis step, and this environment is improper for RNase to have DNA liberated of RNA and even for DNase to degrade the DNA. Therefore, the complete removal of RNA under RNase influence is achieved when RNase is added after DNA extraction, which gives optimal quality with any protocols. Similarly, DNA contamination in an isolated RNA is degraded by DNase to obtain high-quality RNA. Our protocol is the protocol of choice in terms of simplicity, recovery time, environmental safety, amount, purity, PCR and RT-PCR applicability.