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

Kousuke Niwa; Akinobu Oribe; Hidemasa Okumura; Masahiro Shimono; Kenkichi Nagai; Toshikazu Hirota; Hiroshi Yasue; Mitsuo Kawase

doi:10.1016/j.ab.2014.07.010

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

Kousuke Niwa, Akinobu Oribe, Hidemasa Okumura, Masahiro Shimono, Kenkichi Nagai, Toshikazu Hirota, Hiroshi Yasue, Mitsuo Kawase

MEN - Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

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.

Original language	English
Pages (from-to)	12-16
Number of pages	5
Journal	Analytical Biochemistry
Volume	464
DOIs	https://doi.org/10.1016/j.ab.2014.07.010
Publication status	Published - 2014 Nov 1

Keywords

DNA microarray
Fluorescence label
Hybridization
Non-nucleic acid spacer
Single-stranded DNA tag

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10.1016/j.ab.2014.07.010

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title = "Tag/hybridization-based sensitive detection of polymerase chain reaction products",

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.",

keywords = "DNA microarray, Fluorescence label, Hybridization, Non-nucleic acid spacer, Single-stranded DNA tag",

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T1 - Tag/hybridization-based sensitive detection of polymerase chain reaction products

AU - Niwa, Kousuke

AU - Oribe, Akinobu

AU - Okumura, Hidemasa

AU - Shimono, Masahiro

AU - Nagai, Kenkichi

AU - Hirota, Toshikazu

AU - Yasue, Hiroshi

AU - Kawase, Mitsuo

PY - 2014/11/1

Y1 - 2014/11/1

N2 - 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.

AB - 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.

KW - DNA microarray

KW - Fluorescence label

KW - Hybridization

KW - Non-nucleic acid spacer

KW - Single-stranded DNA tag

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

ER -

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

Abstract

Keywords

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