PMID- 18471308
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20110714
LR  - 20230510
IS  - 1755-8166 (Electronic)
IS  - 1755-8166 (Linking)
VI  - 1
DP  - 2008 Mar 26
TI  - Direct fluorescent labelling of clones by DOP PCR.
PG  - 3
LID - 10.1186/1755-8166-1-3 [doi]
AB  - BACKGROUND: Array Comparative Genomic Hybridisation (array CGH) is a powerful 
      technique for the analysis of constitutional chromosomal anomalies. Chromosomal 
      duplications or deletions detected by array CGH need subsequently to be validated 
      by other methods. One method of validation is Fluorescence in situ Hybridisation 
      (FISH). Traditionally, fluorophores or hapten labelling is performed by nick 
      translation or random prime labelling of purified Bacterial Artificial Chromosome 
      (BAC) products. However, since the array targets have been generated from 
      Degenerate Oligonucleotide Primed (DOP) amplified BAC clones, we aimed to use 
      these DOP amplified BAC clones as the basis of an automated FISH labelling 
      protocol. Unfortunately, labelling of DOP amplified BAC clones by traditional 
      labelling methods resulted in high levels of background. RESULTS: We designed an 
      improved labelling method, by means of degenerate oligonucleotides that resulted 
      in optimal FISH probes with low background. CONCLUSION: We generated an improved 
      labelling method for FISH which enables the rapid generation of FISH probes 
      without the need for isolating BAC DNA. We labelled about 900 clones with this 
      method with a success rate of 97%.
FAU - Backx, Liesbeth
AU  - Backx L
AD  - Center for Human Genetics, University Hospital Leuven, Herestraat 49, B-3000 
      Leuven, Belgium. Liesbeth.backx@med.kuleuven.be.
FAU - Thoelen, Reinhilde
AU  - Thoelen R
FAU - Van Esch, Hilde
AU  - Van Esch H
FAU - Vermeesch, Joris R
AU  - Vermeesch JR
LA  - eng
GR  - WT_/Wellcome Trust/United Kingdom
PT  - Journal Article
DEP - 20080326
PL  - England
TA  - Mol Cytogenet
JT  - Molecular cytogenetics
JID - 101317942
PMC - PMC2375879
EDAT- 2008/05/13 09:00
MHDA- 2008/05/13 09:01
PMCR- 2008/03/26
CRDT- 2008/05/13 09:00
PHST- 2007/10/31 00:00 [received]
PHST- 2008/03/26 00:00 [accepted]
PHST- 2008/05/13 09:00 [pubmed]
PHST- 2008/05/13 09:01 [medline]
PHST- 2008/05/13 09:00 [entrez]
PHST- 2008/03/26 00:00 [pmc-release]
AID - 1755-8166-1-3 [pii]
AID - 10.1186/1755-8166-1-3 [doi]
PST - epublish
SO  - Mol Cytogenet. 2008 Mar 26;1:3. doi: 10.1186/1755-8166-1-3.