Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension

PCR-driven overlap extension is a simple and versatile tool in performing site-directed mutagenesis. This technique contains two stages of PCR. The first stage is the initial PCR to generate mutated gene segments that overlapped, which are then used as a DNA template in the second stage PCR to produ...

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Main Authors: Karim, Mohd Shahrul Nizwanshah, Abdullah, Siti Nor Akmar
Format: Conference or Workshop Item
Language:English
Published: Institute of Plantation Studies, Universiti Putra Malaysia 2017
Online Access:http://psasir.upm.edu.my/id/eprint/58843/1/Poster_Paper_21.pdf
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author Karim, Mohd Shahrul Nizwanshah
Abdullah, Siti Nor Akmar
author_facet Karim, Mohd Shahrul Nizwanshah
Abdullah, Siti Nor Akmar
author_sort Karim, Mohd Shahrul Nizwanshah
collection UPM
description PCR-driven overlap extension is a simple and versatile tool in performing site-directed mutagenesis. This technique contains two stages of PCR. The first stage is the initial PCR to generate mutated gene segments that overlapped, which are then used as a DNA template in the second stage PCR to produce full length product with the site-directed mutation. The internal primer serves to introduce nucleotide substitution for site-directed mutagenesis by complementary 3’ ends on the intermediate segments. The full-length product amplied by the flanking primers in the second stage PCR includes the restriction enzyme sites for inserting the product into an expression vector for cloning purposes.
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spelling upm.eprints-588432018-02-07T03:40:32Z http://psasir.upm.edu.my/id/eprint/58843/ Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension Karim, Mohd Shahrul Nizwanshah Abdullah, Siti Nor Akmar PCR-driven overlap extension is a simple and versatile tool in performing site-directed mutagenesis. This technique contains two stages of PCR. The first stage is the initial PCR to generate mutated gene segments that overlapped, which are then used as a DNA template in the second stage PCR to produce full length product with the site-directed mutation. The internal primer serves to introduce nucleotide substitution for site-directed mutagenesis by complementary 3’ ends on the intermediate segments. The full-length product amplied by the flanking primers in the second stage PCR includes the restriction enzyme sites for inserting the product into an expression vector for cloning purposes. Institute of Plantation Studies, Universiti Putra Malaysia 2017 Conference or Workshop Item PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/58843/1/Poster_Paper_21.pdf Karim, Mohd Shahrul Nizwanshah and Abdullah, Siti Nor Akmar (2017) Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension. In: International Conference on Big Data Applications in Agriculture (ICBAA2017), 5-6 Dec. 2017, Auditorium Putra, TNCPI Building, Universiti Putra Malaysia. (pp. 191-195). http://spel2.upm.edu.my/webupm/upload/dokumen/penerbitan/20180102001241ICBAA2017_Poster_Paper_21.pdf
spellingShingle Karim, Mohd Shahrul Nizwanshah
Abdullah, Siti Nor Akmar
Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension
title Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension
title_full Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension
title_fullStr Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension
title_full_unstemmed Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension
title_short Mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene (HGGT) by PCR-driven overlap extension
title_sort mutagenesis of the promoter of the oil palm homogentisate geranylgeranyl transferase gene hggt by pcr driven overlap extension
url http://psasir.upm.edu.my/id/eprint/58843/1/Poster_Paper_21.pdf
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AT abdullahsitinorakmar mutagenesisofthepromoteroftheoilpalmhomogentisategeranylgeranyltransferasegenehggtbypcrdrivenoverlapextension