doi: 10.1016/0921-8777(94)90018-3.
Efficient synthesis of 32P-labeled single-stranded DNA probes using linear PCR; application of the method for analysis of strand-specific DNA repair
Affiliations
- PMID: 7521000
- DOI: 10.1016/0921-8777(94)90018-3
Item in Clipboard
Efficient synthesis of 32P-labeled single-stranded DNA probes using linear PCR; application of the method for analysis of strand-specific DNA repair
Mutat Res.
1994 Sep.
Abstract
We have developed a rapid method to synthesize radioactively labeled single-stranded DNA probes suitable for strand-specific analysis of single copy genes on Southern blot. Linear PCR with 10 microCi alpha 32P-dATP (3000 Ci/mmol) as the only dATP source enabled us to generate strand-specific DNA probes with high specific activity. The probes synthesized by this method have higher specific activities and the same strand specificity compared to the end-labeled single-stranded DNA probes obtained from single-stranded M13mp18/19 vectors. Application of the method for strand-specific analysis of ultraviolet-induced DNA lesions in defined DNA sequences significantly improved the hybridization signal.
Similar articles
-
"Run-off" synthesis and application of defined single-stranded DNA hybridization probes.Anal Biochem. 1990 Feb 15;185(1):164-9. doi: 10.1016/0003-2697(90)90274-d. Anal Biochem. 1990. PMID: 1693048
-
Generation of digoxigenin-labelled double-stranded and single-stranded probes using the polymerase chain reaction.Mol Cell Probes. 1992 Jun;6(3):193-200. doi: 10.1016/0890-8508(92)90016-q. Mol Cell Probes. 1992. PMID: 1406727
-
'Run-off' polymerization with digoxigenin labelled nucleotides creates highly sensitive and strand specific DNA hybridization probes: synthesis and application.Mol Cell Probes. 1992 Apr;6(2):107-14. doi: 10.1016/0890-8508(92)90054-2. Mol Cell Probes. 1992. PMID: 1381047
-
[Polymerase chain reaction, cold probes and clinical diagnosis].Sante. 1994 Jan-Feb;4(1):43-52. Sante. 1994. PMID: 7909267 Review. French.
-
DNA probes: applications of the principles of nucleic acid hybridization.Crit Rev Biochem Mol Biol. 1991;26(3-4):227-59. doi: 10.3109/10409239109114069. Crit Rev Biochem Mol Biol. 1991. PMID: 1718662 Review.
Cited by
-
The 3' processing factor CstF functions in the DNA repair response.Nucleic Acids Res. 2008 Apr;36(6):1792-804. doi: 10.1093/nar/gkn005. Epub 2008 Feb 5. Nucleic Acids Res. 2008. PMID: 18252771 Free PMC article.
-
RNase-sensitive DNA modification(s) initiates S. pombe mating-type switching.Genes Dev. 2004 Apr 1;18(7):794-804. doi: 10.1101/gad.289404. Epub 2004 Apr 1. Genes Dev. 2004. PMID: 15059961 Free PMC article.
-
Nucleotide excision repair in rat male germ cells: low level of repair in intact cells contrasts with high dual incision activity in vitro.Nucleic Acids Res. 2001 Apr 15;29(8):1791-800. doi: 10.1093/nar/29.8.1791. Nucleic Acids Res. 2001. PMID: 11292852 Free PMC article.
-
Transcription-coupled repair removes both cyclobutane pyrimidine dimers and 6-4 photoproducts with equal efficiency and in a sequential way from transcribed DNA in xeroderma pigmentosum group C fibroblasts.EMBO J. 1995 Jan 16;14(2):360-7. doi: 10.1002/j.1460-2075.1995.tb07010.x. EMBO J. 1995. PMID: 7835346 Free PMC article.
-
Architecture of the replication fork stalled at the 3' end of yeast ribosomal genes.Mol Cell Biol. 2000 Aug;20(15):5777-87. doi: 10.1128/MCB.20.15.5777-5787.2000. Mol Cell Biol. 2000. PMID: 10891513 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
