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每个实验室都能采用的自制DNA梯状条带的简易方法。

Easy method for production of a home-made DNA ladder in every laboratory.

作者信息

Abbasian Mahdi, Seyedi Hadieh Alsadat Eslampanah, Boroujeni Zahra Khalili, Mofid Mohammad Reza

机构信息

Bioinformatics Research Center, Department of Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Adv Biomed Res. 2015 Mar 25;4:70. doi: 10.4103/2277-9175.153894. eCollection 2015.

DOI:10.4103/2277-9175.153894
PMID:25878995
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4386206/
Abstract

BACKGROUND

Molecular DNA markers are one of the essential tools in molecular biology labs with varied applications. In the present study, we suggest an efficient and available strategy to produce molecular size marker in routine laboratories.

MATERIALS AND METHODS

To achieve the desired sizes of DNA fragments, we recruited PCR and bioinformatics techniques to synthesize 14 DNA fragments ranging from 100 to 3000 bp.

RESULTS

Holistic analysis of different parameters in primers design resulted in amplification of fragments in just one PCR program without any by-product and purification step. Our applied method enables researchers to modify amplified DNA fragments by wide range of chemical modifications toward varied applications.

CONCLUSION

Method of home-made DNA ladder production by available ingredients and routine techniques reported in this study can be used in common laboratories for different applications.

摘要

背景

分子DNA标记是分子生物学实验室中具有多种应用的重要工具之一。在本研究中,我们提出了一种在常规实验室中生产分子大小标记的有效且可行的策略。

材料与方法

为了获得所需大小的DNA片段,我们采用PCR和生物信息学技术合成了14个大小从100到3000 bp不等的DNA片段。

结果

对引物设计中不同参数的整体分析使得在仅一个PCR程序中就能扩增出片段,无需任何副产物和纯化步骤。我们应用的方法使研究人员能够通过广泛的化学修饰来修饰扩增的DNA片段,以用于各种不同的应用。

结论

本研究报道的利用现有成分和常规技术自制DNA梯度的方法可在普通实验室用于不同的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b20/4386206/128e13da7301/ABR-4-70-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b20/4386206/bd236e4b6db7/ABR-4-70-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b20/4386206/128e13da7301/ABR-4-70-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b20/4386206/bd236e4b6db7/ABR-4-70-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b20/4386206/128e13da7301/ABR-4-70-g004.jpg

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本文引用的文献

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Optimization of Taq DNA polymerase enzyme expression in Escherichia coli.大肠杆菌中Taq DNA聚合酶表达的优化
Adv Biomed Res. 2012;1:82. doi: 10.4103/2277-9175.103004. Epub 2012 Oct 31.
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Separation and Size Determination of DNA over a 10-,200 kbp Range.
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Preparation of DNA ladder based on multiplex PCR technique.
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Laboratory production of 100 base pair DNA molecular weight markers.100个碱基对DNA分子量标记物的实验室制备
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Click chemistry to construct fluorescent oligonucleotides for DNA sequencing.用于DNA测序的点击化学构建荧光寡核苷酸。
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