PMID- 10415338
OWN - NLM
STAT- MEDLINE
DCOM- 19990909
LR  - 20190707
IS  - 0378-1119 (Print)
IS  - 0378-1119 (Linking)
VI  - 235
IP  - 1-2
DP  - 1999 Jul 22
TI  - Context-dependent Taq-polymerase-mediated nucleotide alterations, as revealed by 
      direct sequencing of the ZNF189 gene: implications for mutation detection.
PG  - 103-9
AB  - We have recently reported on the genetic organisation of a novel Kruppel-like
      zinc finger, ZNF189, located to 9q22-q31. In that study we found no mutations in 
      the coding sequence when using ZNF189 as a candidate gene for sporadic basal cell
      cancer and squamous cell cancer. Here, by direct sequencing of the proximal
      promotor of ZNF189, mutations were found to appear in a small hot-spot region in 
      over 50% of analysed tumour samples, the majority being G to A substitutions. The
      hot-spot region spans a 24bp G-rich region. Repeated analyses of the original
      sample lysates fail to confirm each of these mutations; and frequently new
      mutations appear at neighbouring positions. Subsequent analysis with serial
      dilutions of genomic DNA and a cosmid harbouring the wild-type ZNF189 gene
      demonstrate that these sequence-specific alterations arise in the outer
      PCR-amplification when 50 copies or less of template are used. Although the
      mechanism of how these context-specific alterations arise is not proven, the
      results demonstrate a previously unreported type of PCR-mediated
      sequence-specific alteration that easily could have been interpreted as being of 
      clinical relevance. The phenomena observed show that mutations detected by direct
      sequencing can be caused by PCR-introduced alterations. Consequently, this should
      be of general caution in mutation analysis of disease gene candidates when using 
      small amounts of template, such as microdissected biopsies.
FAU - Odeberg, J
AU  - Odeberg J
AD  - Department of Biotechnology, KTH Royal Institute of Technology, Teknikringen 30, 
      S-10044, Stockholm, Sweden.
FAU - Ahmadian, A
AU  - Ahmadian A
FAU - Williams, C
AU  - Williams C
FAU - Uhlen, M
AU  - Uhlen M
FAU - Ponten, F
AU  - Ponten F
FAU - Lundeberg, J
AU  - Lundeberg J
LA  - eng
SI  - GENBANK/AF025772
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Gene
JT  - Gene
JID - 7706761
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Nucleotides)
RN  - 0 (ZNF189 protein, human)
RN  - EC 2.7.7.- (Taq Polymerase)
SB  - IM
MH  - Artifacts
MH  - Base Sequence
MH  - Carcinoma, Basal Cell/genetics
MH  - Carcinoma, Squamous Cell/genetics
MH  - Chromosomes, Human, Pair 9/genetics
MH  - *DNA Mutational Analysis/methods
MH  - DNA-Binding Proteins/*genetics
MH  - Humans
MH  - Introns/genetics
MH  - Loss of Heterozygosity
MH  - Molecular Sequence Data
MH  - *Mutation
MH  - Nucleotides/*genetics
MH  - Polymerase Chain Reaction
MH  - Promoter Regions, Genetic/genetics
MH  - Taq Polymerase/*metabolism
MH  - Templates, Genetic
EDAT- 1999/07/23 00:00
MHDA- 1999/07/23 00:01
CRDT- 1999/07/23 00:00
PHST- 1999/07/23 00:00 [pubmed]
PHST- 1999/07/23 00:01 [medline]
PHST- 1999/07/23 00:00 [entrez]
AID - S0378-1119(99)00205-X [pii]
AID - 10.1016/s0378-1119(99)00205-x [doi]
PST - ppublish
SO  - Gene. 1999 Jul 22;235(1-2):103-9. doi: 10.1016/s0378-1119(99)00205-x.