Characterization of β-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA

Corinne Wong, Carol E. Dowling, Randall K. Saiki, Russell G. Higuchi, Henry A. Erlich, Haig H. Kazazian

Research output: Contribution to journalArticlepeer-review

Abstract

Direct sequencing of specific regions of genomic DNA1 became feasible with the invention of the polymerase chain reaction (PCR) which permits amplification of specific regions of DNA2-5. Recently, human mitochondria! DNA was amplified and directly sequenced6. Using a thermostable DNA polymerase of T. aquaticus (Saiki, R. K. et al., manuscript in preparation) in the PCR, we have applied a combination of PCR and direct sequence analysis of the amplified product to a human single-copy gene. We studied the genomic DNA of five patients with β-thalassaemia whose mutant alleles were uncharacterized, and found two previously undescribed mutations, along with three known alleles. One new allele is a frameshift at codons 106-107 and the other is an A-C transversion at the cap site (+1) of the β-globin gene. This latter is the first natural mutation observed at the cap site and it occurs in a gene which is poorly expressed.

Original languageEnglish (US)
Pages (from-to)384-386
Number of pages3
JournalNature
Volume330
Issue number6146
DOIs
StatePublished - 1987

ASJC Scopus subject areas

  • General

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