Real time RT-PCR as a tool for quantitative Hevea gene expression studies

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Journal of Rubber Research (JRR)

2011

Real time RT-PCR as a tool for quantitative Hevea gene expression studies

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File	Description	Size	Format
ATTACHMENT01	azlina_2011_real_time_RT-PCR	1 MB	Adobe Acrobat PDF
MARCXML	Real time RT-PCR as a tool for quantitative Hevea gene expression studies	2 KB	XML Document

Title: Real time RT-PCR as a tool for quantitative Hevea gene expression studies
Creator: Azlina Bahari
Creator: Chow KS
Contributor: Hafsah bt. Ahmad
Contributor: Nor Faezah Bt. Ismail
Subject: PRRIM
Subject: Hevea
Date: 2011-04-5 |2011-04-5 |2001– |2011 | | | | | | |
Publisher: Kuala Lumpur : Malaysian Rubber Board
Description: We describe a real time quantitative RT-PCR (qRT-PCR) approach to measure gene expression of selected latex genes with SYBR Green I as the reporter dye. In this study, the target genes were the small rubber particle protein (SRPP) and farnesyl diphosphate synthas (FPPS) while the reference was the 18S ribosomal RNA (rRNA) gene. As a first requirement, amplification efficiencies of primer pairs specific to target and reference genes were determined by two-step RT-PCR. In the RT (reverse transcription) step, single-stranded cDNA was synthesised from latex total RNA. Then, ten-fold serial dilutions were amplified with gene-specific primers to generate cycle threshold (Ct) values for constructing a cDNA standard curve for each gene. Subsequently, one-step qRT-PCR reactions were set up to generate Ct values for the target and reference genes from the same RNA sample. A relative quantification method which employs an efficiency-compensated comparative Ct approach was used to calculate target gene transcript level relative to the 18S rRNA gene.

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collection(s): 2011
Identifier: 15111768
Identifier: valet-20110405-121311
Identifier: vital1:78643
Format: Adobe Acrobat PDF
Coverage: 2011
Language: EN
Relation: 14,1,-,51,64
Relation: J. Rubb. Res. 2011 Vol.14 (1) p.51-64
Rights: No|Yes
Type: Article
Source: JRR
Source: RRIM/LGM

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Azlina Bahari, Chow KS. Kuala Lumpur : Malaysian Rubber Board; 2011-04-5 |2011-04-5 |2001– |2011 | | | | | | |. Real time RT-PCR as a tool for quantitative Hevea gene expression studies.

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2011

Real time RT-PCR as a tool for quantitative Hevea gene expression studies

VITAL Repository 5.2