Fast extraction, amplification and analysis of genes from human blood.
In order to shorten the time spent on the sample preparation for gene analysis, a novel method was proposed through the combination of fast DNA extraction and purification by Generation capture disk, amplification by capillary polymerase chain reaction, and confirmation of amplification products by microchip electrophoresis. With this method, 3 microL blood was enough to obtain adequate target fragments in human genes. Under the optimal conditions in each step, the sample preparation for eight fragments in beta-globin gene and four fragments in ras gene could be finished within 20 min. Since all the experiments were performed on commercial instruments, this method showed a wide range of applicability. In addition, other advantages such as fast speed and low consumption of samples were demonstrated. All these merits proved that such a combination method was of great potential for the clinical diagnostics.
Zhang
Lihua
L
Department of Molecular and Pharmaceutical Biotechnology, Graduate School of Pharmaceutical Sciences, The University of Tokushima, CREST, Japan Science and Technology Corporation (JST), Tokushima 770-8505, Japan.
Dang
Fuquan
F
Kaji
Noritada
N
Baba
Yoshinobu
Y
D001483:Base Sequence; D001769:Blood; D017931:DNA Primers; D046209:Electrophoresis, Microchip; D011905:Genes, ras; D005914:Globins / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D005914:Globins