利用微波炉和干浴加热装置在水稻中分离与 PCR 兼容的 miniprep DNA

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-07-02 DOI:10.1007/s40415-024-01023-w
Dip Pal, Narottam Dey
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引用次数: 0

摘要

植物分子生物学家需要一种有效、快速的微型 DNA 分离方案,用两种现代加热设备(微波炉和干浴)代替传统的水浴,后者不仅耗时,有时还会造成微生物污染。虽然当今大多数现代化实验室都配备了这两种最新的加热设备,但使用这两种设备分离高质量、理想数量的植物基因组 DNA 的报道却非常有限。为了满足这一需求,本研究对早先采用的 DNA 分离方案进行了重新标准化,在不同的水稻组织系苗、叶、茎和根中加入微波和干浴,特别是加热的持续时间和温度,然后通过标准统计分析,将分离出的基因组 DNA 的产量和质量与常用方案进行比较。在从不同植物组织中分离基因组 DNA 时,配对 t 检验无法检测出早期标准化方案与两种新标准化方案(采用微波和干浴)之间在质量和数量上的显著差异。分离出的基因组 DNA 经 PCR 扩增后,通过琼脂糖凝胶电泳进行分辨,以确认其与 PCR 的兼容性。
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PCR compatible miniprep DNA isolation in rice using microwave and dry bath based heating devices

An effective and fast miniprep DNA protocol is on demand among plant molecular biologist where two moderns heating devices (microwave and dry bath) may be adopted in place of conventional water bath which is time consuming and sometimes result in microbial contamination. Though most of the present-day modern laboratories are equipped with these two recent heating devices, very limited reports are available using these two devices for isolation of high-quality plant genomic DNA with desirable quantity. Addressing this demand, in present study an earlier adopted DNA isolation protocol was re-standardized on different rice tissue lines seedlings, leaves, stem and root with the inclusion of microwave and dry bath particularly for the duration and applied temperature during heating followed by comparing with popularly used protocol for yield and quality of the isolated genomic DNA through standard statistical analysis. Pair- wise t test unable to detect any significant differences (for both quality and quantity) between the earlier standardized protocol with the two newly standardized protocol (adopted with microwave and dry bath) for the isolated genomic DNA from the different plant tissues included. The isolated genomic DNA was amplified through PCR followed by resolving through agarose gel electrophoresis to confirm their PCR compatibility.

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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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