Suppr超能文献

DNA、RNA及蛋白质提取:过去与现在

DNA, RNA, and protein extraction: the past and the present.

作者信息

Tan Siun Chee, Yiap Beow Chin

机构信息

School of Postgraduate Studies & Research, Division of Pharmacy, International Medical University, No. 126, Jalan 19/155B, Bukit Jalil, 57000 Kuala Lumpur, Malaysia.

出版信息

J Biomed Biotechnol. 2009;2009:574398. doi: 10.1155/2009/574398.

Abstract

Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination.

摘要

DNA、RNA和蛋白质的提取是分子生物学中使用的基本方法。这些生物分子可以从任何生物材料中分离出来,用于后续的下游过程、分析或制备目的。过去,核酸的提取和纯化过程曾经很复杂、耗时、费力,而且总体通量有限。目前,有许多专门的方法可用于提取纯生物分子,如基于溶液和基于柱的方案。随着时间的推移,手动方法肯定有了很大的改进,有各种商业产品可供选择,其中包括完整的试剂盒,包含分离核酸所需的大部分组件,但大多数都需要重复离心步骤,然后根据样本类型去除上清液,并进行额外的机械处理。近年来,为中大型实验室设计的自动化系统需求不断增长。它是劳动密集型手动方法的替代方案。该技术应允许高通量处理样本;生物分子的产量、纯度、可重复性和可扩展性以及检测的速度、准确性和可靠性应达到最大值,同时将交叉污染的风险降至最低。

相似文献

1
DNA, RNA, and protein extraction: the past and the present.
J Biomed Biotechnol. 2009;2009:574398. doi: 10.1155/2009/574398.
2
Reliability and performance of commercial RNA and DNA extraction kits for FFPE tissue cores.
PLoS One. 2017 Jun 22;12(6):e0179732. doi: 10.1371/journal.pone.0179732. eCollection 2017.
3
A column-based rapid method for the simultaneous isolation of DNA, RNA, miRNA and proteins.
Cell Biol Int. 2012 Sep;36(9):779-83. doi: 10.1042/CBI20110342.
5
Planning Implications Related to Sterilization-Sensitive Science Investigations Associated with Mars Sample Return (MSR).
Astrobiology. 2022 Jun;22(S1):S112-S164. doi: 10.1089/AST.2021.0113. Epub 2022 May 19.
6
Simultaneously extracting DNA, RNA, and protein using kits: is sample quantity or quality prejudiced?
Anal Biochem. 2013 Feb 1;433(1):10-8. doi: 10.1016/j.ab.2012.10.006. Epub 2012 Oct 12.
7
A sequential co-extraction method for DNA, RNA and protein recovery from soil for future system-based approaches.
J Microbiol Methods. 2014 Aug;103:118-23. doi: 10.1016/j.mimet.2014.06.004. Epub 2014 Jun 11.
9
Commercially available kits for manual and automatic extraction of nucleic acids from formalin-fixed, paraffin-embedded (FFPE) tissues.
Acta Dermatovenerol Alp Pannonica Adriat. 2015;24(3):47-53. doi: 10.15570/actaapa.2015.12.
10
Automated DNA extraction from pollen in honey.
Food Chem. 2014 Apr 15;149:302-6. doi: 10.1016/j.foodchem.2013.10.129. Epub 2013 Nov 1.

引用本文的文献

1
Next-generation sequencing applications in food science: fundamentals and recent advances.
Front Bioeng Biotechnol. 2025 Aug 20;13:1638957. doi: 10.3389/fbioe.2025.1638957. eCollection 2025.
3
Improved Degradome Sequencing Protocol via Reagent Recycling from sRNAseq Library Preparations.
Int J Mol Sci. 2025 Jul 21;26(14):7020. doi: 10.3390/ijms26147020.
4
Investigation and optimization of DNA isolation efficiency using ferrite-based magnetic nanoparticles.
Biotechnol Rep (Amst). 2025 Jun 30;47:e00904. doi: 10.1016/j.btre.2025.e00904. eCollection 2025 Sep.
5
Interaction of Nanomaterials with Nucleic Acids and Their Applications in Nucleic Acid Analysis.
Int J Biol Sci. 2025 Jun 9;21(9):4051-4068. doi: 10.7150/ijbs.113309. eCollection 2025.
8
The development and applications of circulating tumour cells, circulating tumour DNA and other emerging biomarkers for early cancer detection.
Explor Target Antitumor Ther. 2025 May 13;6:1002314. doi: 10.37349/etat.2025.1002314. eCollection 2025.
9
A novel approach for DNA extraction of white spot syndrome virus detection in penaeid shrimp.
Sci Rep. 2025 May 5;15(1):15636. doi: 10.1038/s41598-025-98536-5.
10
Modulation of the Main Resistance-Associated ABC Transporter's Expression by Plant Flavonol Isorhamnetin.
Pharmaceuticals (Basel). 2025 Mar 28;18(4):494. doi: 10.3390/ph18040494.

本文引用的文献

2
Magnetic particles for the separation and purification of nucleic acids.
Appl Microbiol Biotechnol. 2006 Dec;73(3):495-504. doi: 10.1007/s00253-006-0675-0. Epub 2006 Oct 25.
3
Friedrich Miescher and the discovery of DNA.
Dev Biol. 2005 Feb 15;278(2):274-88. doi: 10.1016/j.ydbio.2004.11.028.
4
Protein extraction from plant tissues.
Methods Mol Biol. 2004;244:21-7. doi: 10.1385/1-59259-655-x:21.
5
Glass bead purification of plasmid template DNA for high throughput sequencing of mammalian genomes.
Nucleic Acids Res. 2002 Apr 1;30(7):e32. doi: 10.1093/nar/30.7.e32.
6
Tech.Sight. Robotic laboratory automation.
Science. 2002 Jan 18;295(5554):517-8. doi: 10.1126/science.295.5554.517.
7
An improved method for Southwestern blotting.
Front Biosci. 1997 Jun 15;2:c9-11. doi: 10.2741/A166.
8
The detection of DNA-binding proteins by protein blotting.
Nucleic Acids Res. 1980 Jan 11;8(1):1-20. doi: 10.1093/nar/8.1.1.
9
A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
Nucleic Acids Res. 1979 Nov 24;7(6):1513-23. doi: 10.1093/nar/7.6.1513.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验