NISCAIR Online Periodicals Repository

NISCAIR ONLINE PERIODICALS REPOSITORY (NOPR)  >
NISCAIR PUBLICATIONS >
Research Journals >
Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.42 [2005] >
IJBB Vol.42(5) [October 2005] >


Title: Analysis of solid-phase allele-specific primer extension characteristics on biochip in combination with modified primers and PicoGreen staining method
Authors: Qin, Shengying
Liu, Jianhua
Keywords: Single nucleotide polymorphism
Allele-specific primer extension
DNA microarray
Extension efficiency and specificity
Genotyping
PicoGreen staining
Issue Date: Oct-2005
Publisher: CSIR
IPC CodeC07H21/04; C12N15/00; C1215/90
Abstract: DNA microarray technology offers potential for future high-throughput variation genotyping. Allele-specific primer extension procedure on microarray has been considered as an efficient method for single nucleotide polymorphism (SNP) genotyping. However, the high cost of the fluorescent-labeled dNTP used for signal detection in this method limits its application. In the present study, we evaluated the characteristics of solid-phase allele-specific primer extension, in terms of specificity and efficiency and demonstrated that compared to liquid-phase reaction, it requires lower annealing temperature, and higher template and Mg²⁺concentrations. The extension efficiency and specificity were though linked, behave diametric during the gradient change of template and Mg²⁺concentrations or annealing temperature. To obtain both optimal signal intensity and specificity, we introduced an artificial mismatched base at the third position from the primer 3′end, which enhanced the specificity significantly. The PicoGreen staining method, which could decrease the cost greatly, was then introduced to replace the fluorescent-labeled dNTP for signal detection.
Page(s): 308-314
ISSN: 0301-1208
Source:IJBB Vol.42(5) [October 2005]

Files in This Item:

File Description SizeFormat
IJBB 42(5) 308-314.pdf940.35 kBAdobe PDFView/Open
 Current Page Visits: 728 
Recommend this item

 

National Knowledge Resources Consortium |  NISCAIR Website |  Contact us |  Feedback

Disclaimer: NISCAIR assumes no responsibility for the statements and opinions advanced by contributors. The editorial staff in its work of examining papers received for publication is helped, in an honorary capacity, by many distinguished engineers and scientists.

CC License Except where otherwise noted, the Articles on this site are licensed under Creative Commons License: CC Attribution-Noncommercial-No Derivative Works 2.5 India

Copyright © 2012 The Council of Scientific and Industrial Research, New Delhi. All rights reserved.

Powered by DSpace Copyright © 2002-2007 MIT and Hewlett-Packard | Compliant to OAI-PMH V 2.0

Home Page Total Visits: 622014 since 06-Feb-2009  Last updated on 13-Nov-2014Webmaster: nopr@niscair.res.in