Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana

OBJECTIVE: The aim of the present study is to optimize the PCR conditions required to amplify the promoter sequence of an amino acid transporter having an AT-rich base composition with a high number of tandem repeats. RESULT: Results show that successful amplification can be achieved by performing a...

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Autores principales: Dhatterwal, Pinky, Mehrotra, Sandhya, Mehrotra, Rajesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Research Note
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5706289/
https://www.ncbi.nlm.nih.gov/pubmed/29183338
http://dx.doi.org/10.1186/s13104-017-2982-1
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author Dhatterwal, Pinky
Mehrotra, Sandhya
Mehrotra, Rajesh
author_facet Dhatterwal, Pinky
Mehrotra, Sandhya
Mehrotra, Rajesh
author_sort Dhatterwal, Pinky
collection PubMed
description OBJECTIVE: The aim of the present study is to optimize the PCR conditions required to amplify the promoter sequence of an amino acid transporter having an AT-rich base composition with a high number of tandem repeats. RESULT: Results show that successful amplification can be achieved by performing a 2-step PCR at a lower extension temperature of 65 °C for an increased extension period of 1.5 min/kb, with MgCl(2) concentration ranging from 2.5 to 3.0 mM. The results also suggest that the DNA concentration of about 25–30 ng/µl was essential to achieve this amplification.
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spelling pubmed-57062892017-12-05 Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana Dhatterwal, Pinky Mehrotra, Sandhya Mehrotra, Rajesh BMC Res Notes Research Note OBJECTIVE: The aim of the present study is to optimize the PCR conditions required to amplify the promoter sequence of an amino acid transporter having an AT-rich base composition with a high number of tandem repeats. RESULT: Results show that successful amplification can be achieved by performing a 2-step PCR at a lower extension temperature of 65 °C for an increased extension period of 1.5 min/kb, with MgCl(2) concentration ranging from 2.5 to 3.0 mM. The results also suggest that the DNA concentration of about 25–30 ng/µl was essential to achieve this amplification. BioMed Central 2017-11-28 /pmc/articles/PMC5706289/ /pubmed/29183338 http://dx.doi.org/10.1186/s13104-017-2982-1 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Note
Dhatterwal, Pinky
Mehrotra, Sandhya
Mehrotra, Rajesh
Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana
title Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana
title_full Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana
title_fullStr Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana
title_full_unstemmed Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana
title_short Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana
title_sort optimization of pcr conditions for amplifying an at-rich amino acid transporter promoter sequence with high number of tandem repeats from arabidopsis thaliana
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5706289/
https://www.ncbi.nlm.nih.gov/pubmed/29183338
http://dx.doi.org/10.1186/s13104-017-2982-1
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