Midlands State University Library

The methylation status of plant genomic DNA influences PCR efficiency (Record no. 169310)

MARC details
000 -LEADER
fixed length control field 02673nam a22002897a 4500
003 - CONTROL NUMBER IDENTIFIER
control field ZW-GwMSU
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20250317102218.0
008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION
fixed length control field 250317b |||||||| |||| 00| 0 eng d
022 ## - INTERNATIONAL STANDARD SERIAL NUMBER
International Standard Serial Number 0176-1617
040 ## - CATALOGING SOURCE
Original cataloging agency MSU
Language of cataloging English
Transcribing agency MSU
Description conventions rda
050 00 - LIBRARY OF CONGRESS CALL NUMBER
Classification number QK711.2 JOU
100 1# - MAIN ENTRY--PERSONAL NAME
Personal name Kiselev, K.V.
Relator term author
245 14 - TITLE STATEMENT
Title The methylation status of plant genomic DNA influences PCR efficiency
Statement of responsibility, etc. created by K.V. Kiselev, A.S. Dubrovina, A.P. Tyunin
264 1# - PRODUCTION, PUBLICATION, DISTRIBUTION, MANUFACTURE, AND COPYRIGHT NOTICE
Place of production, publication, distribution, manufacture Amsterdam:
Name of producer, publisher, distributor, manufacturer Elsevier GmbH,
Date of production, publication, distribution, manufacture, or copyright notice 2015.
336 ## - CONTENT TYPE
Source rdacontent
Content type term text
Content type code txt
337 ## - MEDIA TYPE
Source rdamedia
Media type term unmediated
Media type code n
338 ## - CARRIER TYPE
Source rdacarrier
Carrier type term volume
Carrier type code nc
440 ## - SERIES STATEMENT/ADDED ENTRY--TITLE
Title Journal of Plant Physiology,
Volume/sequential designation Volume 175
520 3# - SUMMARY, ETC.
Summary, etc. During the polymerase chain reaction (PCR), which is a versatile and widely used method, certain DNA sequences are rapidly amplified through thermocycling. Although there are numerous protocols of PCR optimization for different applications, little is known about the effect of DNA modifications, such as DNA methylation, on PCR efficiency. Recent studies show that cytosine methylation alters DNA mechanical properties and suggest that DNA methylation may directly or indirectly influence the effectiveness of DNA amplification during PCR. In the present study, using plant DNA, we found that highly methylated plant DNA genomic regions were amplified with lower efficiencies compared to that for the regions methylated at a lower level. The correlation was observed when amplifying stilbene synthase (STS1, STS10) genes of Vitis amurensis, the Actin2 gene of Arabidopsis thaliana, the internal transcribed spacer (AtITS), and tRNAPro of A. thaliana. The level of DNA methylation within the analyzed DNA regions has been analyzed with bisulfite sequencing. The obtained data show that efficient PCRs of highly methylated plant DNA regions can be hampered. Proteinase K treatment of the plant DNA prior to PCR and using HotTaq DNA polymerase improved amplification of the highly methylated plant DNA regions. We suggest that increased DNA denaturation temperatures of the highly methylated DNA and contamination with DNA-binding proteins contribute to the hampered PCR amplification of highly methylated DNA. The data show that it is necessary to use current DNA purification protocols and commercial kits with caution to ensure appropriate PCR product yield and prevent bias toward unmethylated DNA amplification in PCRs.
650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element Arabidopsis thaliana
650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element HotTaq DNA polymerase
650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element Plant DNA extraction
700 1# - ADDED ENTRY--PERSONAL NAME
Personal name Dubrovina, A.S.
Relator term co-author
700 1# - ADDED ENTRY--PERSONAL NAME
Personal name Tyunin, A.P.
Relator term co-author
856 ## - ELECTRONIC LOCATION AND ACCESS
Uniform Resource Identifier https://doi.org/10.1016/j.jplph.2014.10.017
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Source of classification or shelving scheme Library of Congress Classification
Koha item type Journal Article
Holdings
Withdrawn status Lost status Source of classification or shelving scheme Damaged status Not for loan Home library Current library Shelving location Date acquired Serial Enumeration / chronology Total Checkouts Full call number Date last seen Price effective from Koha item type Public note
    Library of Congress Classification     Main Library Main Library - Special Collections 02/10/2015 Vol. 175 (pages59-67)   QK711.2 JOU 17/03/2025 17/03/2025 Journal Article For in house use only