Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Lan, Vo Thi Thuong, Loan, Pham Thi Thanh, Duong, Pham Anh Thuy, Thanh, Le Thi, Ha, Ngo Thi, Thuan, Ta Bich
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2012
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
_version_ 1782227266942533632
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
work_keys_str_mv AT lanvothithuong straightforwardprocedureforlaboratoryproductionofdnaladder
AT loanphamthithanh straightforwardprocedureforlaboratoryproductionofdnaladder
AT duongphamanhthuy straightforwardprocedureforlaboratoryproductionofdnaladder
AT thanhlethi straightforwardprocedureforlaboratoryproductionofdnaladder
AT hangothi straightforwardprocedureforlaboratoryproductionofdnaladder
AT thuantabich straightforwardprocedureforlaboratoryproductionofdnaladder