2003
DOI: 10.1590/s0100-879x2003001100004
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Screening for mutations in human alpha-globin genes by nonradioactive single-strand conformation polymorphism

Abstract: Point mutations and small insertions or deletions in the human α-globin genes may produce α-chain structural variants and α-thalassemia. Mutations can be detected either by direct DNA sequencing or by screening methods, which select the mutated exon for sequencing. Although small (about 1 kb, 3 exons and 2 introns), the α-globin genes are duplicate (α 2 and α 1 ) and highy G-C rich, which makes them difficult to denature, reducing sequencing efficiency and causing frequent artifacts. We modified some condition… Show more

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Cited by 6 publications
(3 citation statements)
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References 12 publications
(9 reference statements)
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“…Several techniques based on PCR amplification of normal and affected chromosomes (26 -28 ) have been developed to more rapidly identify globin gene mutations. These techniques include single-strand conformation polymorphism analysis, denaturing gradient gel electrophoresis (29,30 ), direct sequencing, amplification refractory mutation system PCR (31 ), reverse dot-blot analysis (32 ), and Gap-PCR, which is based on the multiplex amplification of junctional segments of several different breakpoints (33)(34)(35)(36). The latter technique enables screening and diagnosis of several common deletions in a single test.…”
mentioning
confidence: 99%
“…Several techniques based on PCR amplification of normal and affected chromosomes (26 -28 ) have been developed to more rapidly identify globin gene mutations. These techniques include single-strand conformation polymorphism analysis, denaturing gradient gel electrophoresis (29,30 ), direct sequencing, amplification refractory mutation system PCR (31 ), reverse dot-blot analysis (32 ), and Gap-PCR, which is based on the multiplex amplification of junctional segments of several different breakpoints (33)(34)(35)(36). The latter technique enables screening and diagnosis of several common deletions in a single test.…”
mentioning
confidence: 99%
“…Although SSCP does not enjoy wide applicability in globin gene genetic screening, as in a variety of other genomic loci [Garinis et al, 2005], a number of SSCP-based mutation-screening strategies have been described for the HBA2/HBA1 [Harteveld et al, 1996, Jorge et al, 2003, HBB [Takahashi-Fujii et al, 1994, Gupta andAgarwal, 2003], and HBE1 genes (Papachatzopoulou et al, in press). However, careful adjustment of the experimental conditions is required in order to obtain reproducible results between different runs, particularly for the purposes of prenatal diagnosis.…”
Section: Methodology Overview: Symphony Of Athousandmentioning
confidence: 99%
“…The semi-automated electrophoresis unit is an automated and cost-effective device for gene analysis. The device works with precast polyacrylamide gels, as well as semidry buffer strips, eliminating the need for time-wasting liquid handling (Custodio and Carmo-Fonseca 2001;Salazar et al 2002;Jorge et al 2003).…”
Section: Introductionmentioning
confidence: 99%