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Straightforward Procedure for Laboratory Production of DNA Ladder
DNA ladder is commonly used to determine the size of DNA fragments by electrophoresis in routine molecular biology laboratories. In this study, we report a new procedure to prepare a DNA ladder that consists of 10 fragments from 100 to 1000 bp. This protocol is a combination of routinely employed me...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306978/ https://www.ncbi.nlm.nih.gov/pubmed/22496965 http://dx.doi.org/10.1155/2012/254630 |
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author | Lan, Vo Thi Thuong Loan, Pham Thi Thanh Duong, Pham Anh Thuy Thanh, Le Thi Ha, Ngo Thi Thuan, Ta Bich |
author_facet | Lan, Vo Thi Thuong Loan, Pham Thi Thanh Duong, Pham Anh Thuy Thanh, Le Thi Ha, Ngo Thi Thuan, Ta Bich |
author_sort | Lan, Vo Thi Thuong |
collection | PubMed |
description | DNA ladder is commonly used to determine the size of DNA fragments by electrophoresis in routine molecular biology laboratories. In this study, we report a new procedure to prepare a DNA ladder that consists of 10 fragments from 100 to 1000 bp. This protocol is a combination of routinely employed methods: cloning, PCR, and partial digestion with restriction enzymes. DNA fragments of 100 bp with unique restriction site at both ends were self-ligated to create a tandem repeat. Once being cloned, the tandem repeat was rapidly amplified by PCR and partially digested by restriction enzymes to produce a ladder containing multimers of the repeated DNA fragments. Our procedure for production of DNA ladder could be simple, time saving, and inexpensive in comparison with current ones widely used in most laboratories. |
format | Online Article Text |
id | pubmed-3306978 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-33069782012-04-11 Straightforward Procedure for Laboratory Production of DNA Ladder Lan, Vo Thi Thuong Loan, Pham Thi Thanh Duong, Pham Anh Thuy Thanh, Le Thi Ha, Ngo Thi Thuan, Ta Bich J Nucleic Acids Research Article DNA ladder is commonly used to determine the size of DNA fragments by electrophoresis in routine molecular biology laboratories. In this study, we report a new procedure to prepare a DNA ladder that consists of 10 fragments from 100 to 1000 bp. This protocol is a combination of routinely employed methods: cloning, PCR, and partial digestion with restriction enzymes. DNA fragments of 100 bp with unique restriction site at both ends were self-ligated to create a tandem repeat. Once being cloned, the tandem repeat was rapidly amplified by PCR and partially digested by restriction enzymes to produce a ladder containing multimers of the repeated DNA fragments. Our procedure for production of DNA ladder could be simple, time saving, and inexpensive in comparison with current ones widely used in most laboratories. Hindawi Publishing Corporation 2012 2012-02-12 /pmc/articles/PMC3306978/ /pubmed/22496965 http://dx.doi.org/10.1155/2012/254630 Text en Copyright © 2012 Vo Thi Thuong Lan et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Lan, Vo Thi Thuong Loan, Pham Thi Thanh Duong, Pham Anh Thuy Thanh, Le Thi Ha, Ngo Thi Thuan, Ta Bich Straightforward Procedure for Laboratory Production of DNA Ladder |
title | Straightforward Procedure for Laboratory Production of DNA Ladder |
title_full | Straightforward Procedure for Laboratory Production of DNA Ladder |
title_fullStr | Straightforward Procedure for Laboratory Production of DNA Ladder |
title_full_unstemmed | Straightforward Procedure for Laboratory Production of DNA Ladder |
title_short | Straightforward Procedure for Laboratory Production of DNA Ladder |
title_sort | straightforward procedure for laboratory production of dna ladder |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306978/ https://www.ncbi.nlm.nih.gov/pubmed/22496965 http://dx.doi.org/10.1155/2012/254630 |
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