Exon skipping in the ATM gene in normal individuals: the effect of blood sample storage on RT-PCR analysis

Birrell, G. W., Ramsay, J., Tung, J. J. and Lavin, M. F. (2001) Exon skipping in the ATM gene in normal individuals: the effect of blood sample storage on RT-PCR analysis. Human Mutation, 17 1: 75-76. doi:10.1002/1098-1004(2001)17:1


Author Birrell, G. W.
Ramsay, J.
Tung, J. J.
Lavin, M. F.
Title Exon skipping in the ATM gene in normal individuals: the effect of blood sample storage on RT-PCR analysis
Journal name Human Mutation   Check publisher's open access policy
ISSN 1059-7794
Publication date 2001
DOI 10.1002/1098-1004(2001)17:1
Volume 17
Issue 1
Start page 75
End page 76
Total pages 2
Editor R. G. H. Cotton
Haig H. Kazazian Jr.
Place of publication USA
Publisher John Wiley & Sons, Inc
Collection year 2001
Language eng
Subject C1
270103 Protein Targeting and Signal Transduction
730108 Cancer and related disorders
060108 Protein Trafficking
Formatted abstract
Mutations in ATM, the gene defective in the human genetic disorder ataxia-telangiectasia
(A-T), have been described in A-T patients and in a variety of tumor samples. Most of these
arise due to exon skipping. We developed an RT-PCR based protein truncation test (PTT) to
screen for ATM mutations in breast cancer patients showing adverse response to
radiotherapy. An additional PTT product was evident in the ATM gene in peripheral blood
mononuclear cells (PBMCs) from blood samples that were collected 2 days or more prior to
RNA extraction. Lymphoblastoid cell lines established from the same blood samples showed
no evidence of the additional band. Cloning and sequencing of the additional RT-PCR
product revealed an exact deletion of exon 20 (2639 del 200), pointing to exon skipping. RTPCR
analysis of RNA extracted from freshly prepared PBMCs from 3 normal individuals
showed no evidence of the additional RT-PCR product but when the blood was stored for 2-
3 days prior to RNA extraction the lower molecular weight band was evident in every
sample. DNA sequencing confirmed this to be due to loss of exon 20. These data suggest that
mRNA-based mutation analysis on ATM should be carried out on unstored blood samples to
avoid artificial loss of exons that give rise to apparent mutations. © 2000 Wiley-Liss, Inc.
KEY WORDS: ataxia telangiectasia; ATM; exon skipping; protein truncation test; PTT; sample analysis; artifacts
Keyword Ataxia Telangiectasia
ATM
exon skipping
protein truncation test
PTT
sample analysis
artifacts
Q-Index Code C1

Document type: Journal Article
Collection: School of Medicine Publications
 
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Created: Tue, 14 Aug 2007, 15:58:03 EST