Title
Use of the SSCP technique to detect point mutations on human mtDNA
Other title
Uso de la técnica SSCP para detectar mutaciones puntuales del AND mitocondrial humano
Date Issued
01 January 2005
Access level
metadata only access
Resource Type
journal article
Publisher(s)
Facultad de Ciencias Biologicas, Universidad Nacional Mayor de San Marcos
Abstract
We evaluate the use of SSCP (single strand conformational polymorphism), a relatively easy and inexpensive technique for the detection of point mutations with a sensibility around 80% under ideal conditions. To test the technique, we used samples of volunteers whose DNA had been previously characterized for the presence or absence of 5 mitochondrial RFLPs. Optimization of the tests included variations in TBE (1X and 0,5X) and of glycerol concentration (10%, 5% and no glycerol) in polyacrylamide gels. Four out of five RFLPs were detected under the conditions used and could be applied routinely without using restriction enzymes. In addition, the SSCP technique allowed detection of unknown mutations in a 394 bp nucleotide segment of the hypervariable (HVI) region of mtDNA. Differences corresponding to different haplotypes were detected, helping to distinguish groups within the same subtype. Sequencing of two samples of subtype B1 with differential migration on SSCP gels, proved the existence in seven different nucleotides.
Start page
349
End page
358
Volume
12
Issue
3
Language
Spanish
OCDE Knowledge area
Genética humana
Subjects
Scopus EID
2-s2.0-85013952808
Source
Revista Peruana de Biologia
ISSN of the container
15610837
Sponsor(s)
Nuestros resultados muestran la factibilidad de utilizar la técnica SSCP para detectar mu-taciones puntuales conocidas o las desconoci-das, estas últimas, las más numerosas entre todas las mutaciones del genoma y las más difíciles de reconocer.
Sources of information:
Directorio de Producción Científica
Scopus