Denaturing gradient gel electrophoresis and direct sequencing of PCR amplified genomic DNA: a rapid and reliable diagnostic approach to beta thalassaemia
- 1 October 1990
- journal article
- research article
- Published by Wiley in British Journal of Haematology
- Vol. 76 (2) , 269-274
- https://doi.org/10.1111/j.1365-2141.1990.tb07883.x
Abstract
Summary The analysis of polymerase chain reaction (PCR)‐amplified β‐globin DNA with allele‐specific oligonucleotide (ASO) probes reveals a very heterogeneous spectrum of β‐thalassaemia in the Netherlands. However about 20% of the β‐thalassaemia mutations cannot be identified with this approach. The combination of specific amplification of certain regions of the β‐globin gene with denaturing gradient gel electrophoresis (DGGE) allowed us to rapidly localize several of these mutations to specific regions of the gene. which were again amplified and directly sequenced.We believe that the combination of DGGE and the direct sequence determination of PCR amplified genomic DNA represents a valid alternative to the‘ASO probes’approach, especially in countries where a very heterogeneous spectrum of β‐thalassaemia mutations occurs.This publication has 24 references indexed in Scilit:
- Use of denaturing gradient gel electrophoresis to detect point mutations in the factor VIII geneGenomics, 1990
- Identification of the multiple beta-thalassemia mutations by denaturing gradient gel electrophoresis.Journal of Clinical Investigation, 1990
- Mutations in the catalytic domain of human coagulation factor IX: Rapid characterization by direct genomic sequencing of DNA fragments displaying an altered melting behaviorGenomics, 1989
- Frameshift Codon 5 [FSC-5 (−CT)] Thalassemia; a Novel Mutation Detected in a Greek PatientHemoglobin, 1989
- A Further case of β-Thalassemia with an Homozygous T→C Substitution at the Donor Splice Site of the First Intervening Sequence of the β-Globin GeneHemoglobin, 1989
- [31] Detection and localization of single base changes by denaturing gradient gel electrophoresisPublished by Elsevier ,1987
- Analysis of enzymatically amplified β-globin and HLA-DQα DNA with allele-specific oligonucleotide probesNature, 1986
- TECHNOLOGICAL EXAMINATION OF LOW‐FIRED TERRACOTTA STATUES FROM AYIA IRINI, KEAArchaeometry, 1982
- Linkage of β-thalassaemia mutations and β-globin gene polymorphisms with DNA polymorphisms in human β-globin gene clusterNature, 1982
- DNA sequence variants in the Gγ-, Aγ-, δ- and β-globin genes of manCell, 1979