New preparation of PM2 phage DNA and an endonuclease assay for a single-strand break

Sang Oun Jung, Chung Kyoon Auh, Jae Chun Ryu, Joon Kim

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

PM2 is a bacteriophage which has closed circular double-stranded DNA as a genome, which is the sole source for endonuclease assay for a single strand break in the fmol range. Therefore, it is important to isolate PM2 DNA with low control nicks for the endonuclease assay. Usually, the isolation method of phage DNA is to use ultracentrifugation which takes at least 4 days. In this report, a fast and effective method which takes only 2 days was developed to purify DNA using polyethylene glycol (PEG) 8000 and the yields of phage DNA isolated by these two methods were compared. The method using PEG 8000 increased the yield of PM2 DNA from 31.2% to 45.2%, and decreased the nick from 17.1% to 13.1%. Recently, the complete PM2 DNA genome sequence of 10,079 bp was published. The exact number of nucleotides of PM2 DNA is important for the correct enzyme assay which measures nicks generated by an endonuclease. The correct calculation of endonuclease activity of rpS3 for nick-circle assay was performed to measure single-strand breaks in this report.

Original languageEnglish
Pages (from-to)223-229
Number of pages7
JournalAntonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
Volume83
Issue number3
DOIs
Publication statusPublished - 2003 May 28

Fingerprint

Deoxyribonuclease I
Bacteriophages
Endonucleases
DNA
Genome
Ultracentrifugation
Enzyme Assays
Nucleotides

Keywords

  • DNA repair
  • DNA single-strand break
  • Endonuclease
  • Nick circle assay
  • PM2 phage
  • Rps3

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

New preparation of PM2 phage DNA and an endonuclease assay for a single-strand break. / Jung, Sang Oun; Auh, Chung Kyoon; Ryu, Jae Chun; Kim, Joon.

In: Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology, Vol. 83, No. 3, 28.05.2003, p. 223-229.

Research output: Contribution to journalArticle

@article{6eaa38b9c5054abeacfff4c3a65bf4e1,
title = "New preparation of PM2 phage DNA and an endonuclease assay for a single-strand break",
abstract = "PM2 is a bacteriophage which has closed circular double-stranded DNA as a genome, which is the sole source for endonuclease assay for a single strand break in the fmol range. Therefore, it is important to isolate PM2 DNA with low control nicks for the endonuclease assay. Usually, the isolation method of phage DNA is to use ultracentrifugation which takes at least 4 days. In this report, a fast and effective method which takes only 2 days was developed to purify DNA using polyethylene glycol (PEG) 8000 and the yields of phage DNA isolated by these two methods were compared. The method using PEG 8000 increased the yield of PM2 DNA from 31.2{\%} to 45.2{\%}, and decreased the nick from 17.1{\%} to 13.1{\%}. Recently, the complete PM2 DNA genome sequence of 10,079 bp was published. The exact number of nucleotides of PM2 DNA is important for the correct enzyme assay which measures nicks generated by an endonuclease. The correct calculation of endonuclease activity of rpS3 for nick-circle assay was performed to measure single-strand breaks in this report.",
keywords = "DNA repair, DNA single-strand break, Endonuclease, Nick circle assay, PM2 phage, Rps3",
author = "Jung, {Sang Oun} and Auh, {Chung Kyoon} and Ryu, {Jae Chun} and Joon Kim",
year = "2003",
month = "5",
day = "28",
doi = "10.1023/A:1023357432335",
language = "English",
volume = "83",
pages = "223--229",
journal = "Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology",
issn = "0003-6072",
publisher = "Springer Netherlands",
number = "3",

}

TY - JOUR

T1 - New preparation of PM2 phage DNA and an endonuclease assay for a single-strand break

AU - Jung, Sang Oun

AU - Auh, Chung Kyoon

AU - Ryu, Jae Chun

AU - Kim, Joon

PY - 2003/5/28

Y1 - 2003/5/28

N2 - PM2 is a bacteriophage which has closed circular double-stranded DNA as a genome, which is the sole source for endonuclease assay for a single strand break in the fmol range. Therefore, it is important to isolate PM2 DNA with low control nicks for the endonuclease assay. Usually, the isolation method of phage DNA is to use ultracentrifugation which takes at least 4 days. In this report, a fast and effective method which takes only 2 days was developed to purify DNA using polyethylene glycol (PEG) 8000 and the yields of phage DNA isolated by these two methods were compared. The method using PEG 8000 increased the yield of PM2 DNA from 31.2% to 45.2%, and decreased the nick from 17.1% to 13.1%. Recently, the complete PM2 DNA genome sequence of 10,079 bp was published. The exact number of nucleotides of PM2 DNA is important for the correct enzyme assay which measures nicks generated by an endonuclease. The correct calculation of endonuclease activity of rpS3 for nick-circle assay was performed to measure single-strand breaks in this report.

AB - PM2 is a bacteriophage which has closed circular double-stranded DNA as a genome, which is the sole source for endonuclease assay for a single strand break in the fmol range. Therefore, it is important to isolate PM2 DNA with low control nicks for the endonuclease assay. Usually, the isolation method of phage DNA is to use ultracentrifugation which takes at least 4 days. In this report, a fast and effective method which takes only 2 days was developed to purify DNA using polyethylene glycol (PEG) 8000 and the yields of phage DNA isolated by these two methods were compared. The method using PEG 8000 increased the yield of PM2 DNA from 31.2% to 45.2%, and decreased the nick from 17.1% to 13.1%. Recently, the complete PM2 DNA genome sequence of 10,079 bp was published. The exact number of nucleotides of PM2 DNA is important for the correct enzyme assay which measures nicks generated by an endonuclease. The correct calculation of endonuclease activity of rpS3 for nick-circle assay was performed to measure single-strand breaks in this report.

KW - DNA repair

KW - DNA single-strand break

KW - Endonuclease

KW - Nick circle assay

KW - PM2 phage

KW - Rps3

UR - http://www.scopus.com/inward/record.url?scp=0038293476&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038293476&partnerID=8YFLogxK

U2 - 10.1023/A:1023357432335

DO - 10.1023/A:1023357432335

M3 - Article

VL - 83

SP - 223

EP - 229

JO - Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology

JF - Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology

SN - 0003-6072

IS - 3

ER -