HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP
Braz. j. med. biol. res
;
34(7): 867-869, July 2001. ilus
Article
in English
| LILACS
| ID: lil-298678
RESUMO
In the present study we used a simple and reliable method for HLA-DQA1 allele typing based on the single-stranded conformation polymorphism (SSCP) properties of DNA molecules obtained by PCR. The technique consists of PCR amplification of a DNA fragment comprising the second exon of the HLA-DQA1 gene, amplicon denaturation using a low ionic strength solution (LIS), and electrophoresis on a small native polyacrylamide gel, followed by a rapid silver staining procedure. In order to validate the technique and to obtain the allele patterns for the DQA1 gene, 50 cervical samples were typed using this methodology and the commercial Amplitype® HLA DQA1 Amplification and Typing kit. All the alleles detected with the kit were characterized by the LIS-SSCP approach. This procedure proved to be useful for population screening and typing of the DQA1 gene as well as for detecting new alleles or mutations in the donor-recipient molecular matching of HLA class II genes
Full text:
Available
Index:
LILACS (Americas)
Main subject:
HLA-DQ Antigens
/
Alleles
Limits:
Humans
Language:
English
Journal:
Braz. j. med. biol. res
Journal subject:
Biology
/
Medicine
Year:
2001
Type:
Article
Affiliation country:
Argentina
Institution/Affiliation country:
Universidad Nacional de La Plata/AR
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