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Tag/hybridization-based sensitive detection of polymerase chain reaction products

Authors :
Akinobu Oribe
Kousuke Niwa
Okumura Hidemasa
Mitsuo Kawase
Toshikazu Hirota
Masahiro Shimono
Kenkichi Nagai
Hiroshi Yasue
Source :
Analytical Biochemistry. 464:12-16
Publication Year :
2014
Publisher :
Elsevier BV, 2014.

Abstract

The polymerase chain reaction (PCR) is an important technology to amplify a single copy or a few copies of DNA segment in genomic DNAs, visualizing the segment as DNA fragment. Thus, PCR is frequently used in various examinations such as detection of bacteria and fungi in the food industry. Here, we report a simple and sensitive method for detection of PCR products using single-strand tag sequence and hybridization of the tag sequence to the complementary tag sequence immobilized on solid material (STH). The detection sensitivity was found to be at least 50 times higher than electrophoresis/ethidium bromide (EtBr) visualization for approximately a 500-bp fragment and higher than the ordinary hybridization, that is, hybridization of denatured PCR product to probe sequence immobilized on solid material.

Details

ISSN :
00032697
Volume :
464
Database :
OpenAIRE
Journal :
Analytical Biochemistry
Accession number :
edsair.doi.dedup.....47dbb19ab8d9f93c35d57b8449ed4dd5
Full Text :
https://doi.org/10.1016/j.ab.2014.07.010