Randomized terminal linker-dependent PCR: a versatile and sensitive method for detection of DNA damage / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
; (12): 203-208, 2002.
Article
in English
| WPRIM (Western Pacific)
| ID: wpr-264316
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To design and develop a novel, sensitive and versatile method for in vivo foot printing and studies of DNA damage, such as DNA adducts and strand breaks.</p><p><b>METHODS</b>Starting with mammalian genomic DNA, single-stranded products were made by repeated primer extension, these products were ligated to a double-stranded linker having a randomized 3' overhang, and used for PCR. DNA breaks in p53 gene produced by restriction endonuclease AfaI were detected by using this new method followed by Southern hybridization with DIG-labeled probe.</p><p><b>RESULTS</b>This randomized terminal linker-dependent PCR (RDPCR) method could generate band signals many-fold stronger than conventional ligation-mediated PCR (LMPCR), and it was more rapid, convenient and accurate than the terminal transferase-dependent PCR (TDPCR).</p><p><b>CONCLUSION</b>DNA strand breakage can be detected sensitively in the gene level by RDPCR. Any lesion that blocks primer extension should be detectable.</p>
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
DNA Damage
/
Polymerase Chain Reaction
/
Genes, p53
/
Sensitivity and Specificity
/
DNA Primers
/
DNA Adducts
/
Genetics
/
Mammals
/
Methods
Type of study:
Controlled clinical trial
/
Diagnostic study
Limits:
Animals
/
Humans
Language:
English
Journal:
Biomedical and Environmental Sciences
Year:
2002
Document type:
Article