A rapid and cost-effective method for analysis of three common genetic risk factors for thrombosis
作者:
J. Cutler,
M. Mitchell,
K. Greenslade,
M. Smith,
G. Savidge,
期刊:
Blood Coagulation and Fibrinolysis
(OVID Available online 2001)
卷期:
Volume 12,
issue 1
页码: 33-36
ISSN:0957-5235
年代: 2001
出版商: OVID
关键词: thrombosis;risk factors;polymerase chain reaction;Guthrie spots;single-stranded conformational polymorphism
数据来源: OVID
摘要:
A simple, rapid and cost-effective method for the analysis of three of the most widely screened genetic risk factors for thrombosis has been established. The protocol developed uses blood spots stored on filter paper (Guthrie spots) as well as DNA extracted from anticoagulated blood. The use of Guthrie spots taken at birth enables the retrospective study of patients who develop thrombotic complications without necessitating resampling. Following isolation of DNA, conventional fluorescence-labelled polymerase chain reaction (PCR) is performed using a thermostable DNA polymerase. Denatured, single-stranded PCR products are analysed on a semi-automated capillary-based genetic analyser, the data being stored electronically. This sensitive protocol obviates the need for endonuclease digestion and the associated gel running and documentation, and leads to a reduction in the recurrent costs of laboratory consumables.
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