Fast 5-hour determination of angiotensin-converting enzyme genotypes from blood by SDS-PAGE using PhastSystem.
Polymorphism in the angiotensin-converting enzyme (ACE) gene has been shown to correlate with circulating ACE concentrations in plasma, and also to be an independent risk factor in the development of many cardiovascular diseases. However, methods available today for ACE genotyping are still laborious and time-consuming. Here we report a sensitive, simple and non-isotopic procedure with commercially available gels for the identification of ACE insertion/ deletion polymorphism. This technique includes rapid extraction of the DNA by the QIAamp Blood Kit prior to polymerase chain reaction, followed by sodium dodecylsulphate polyacrylamide gel electrophoresis using the PhastSystem (Pharmacia). The procedure can be accomplished in five-hours from drawing the blood samples to the final result.
Huang
X H
XH
Department of Clinical Chemistry, Tampere University Hospital, Finland.
Koivula
T
T
Ilveskoski
E
E
Karhunen
P J
PJ
Rantalaiho
V
V
Wirta
O
O
Pasternack
A
A
Lehtimäki
T
T
D000368:Aged; D000483:Alleles; D004247:DNA / Q000032:analysis / Q000235:genetics; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000201:enzymology / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D007703:Peptidyl-Dipeptidase A / Q000097:blood* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000379:methods; D012967:Sodium Dodecyl Sulfate; D013997:Time Factors
D012967:Sodium Dodecyl Sulfate; D004247:DNA; D007703:Peptidyl-Dipeptidase A