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.

[1]  S. Jacobson,et al.  Integrated system for rapid PCR-based DNA analysis in microfluidic devices. , 2000, Analytical chemistry.

[2]  J. Devaney,et al.  Analysis of DNA restriction fragments and polymerase chain reaction products by capillary electrophoresis. , 1994, Journal of chromatography. A.

[3]  M. Powell,et al.  Detection of the hereditary hemochromatosis gene mutation by real-time fluorescence polymerase chain reaction and peptide nucleic acid clamping. , 1998, Analytical biochemistry.

[4]  P. Wilding,et al.  Microchip module for blood sample preparation and nucleic acid amplification reactions. , 2001, Genome research.

[5]  H. Miller Japanese Pharmaceutical Biotechnology: Perception vs. Reality , 1989, Bio/Technology.

[6]  P E Klapper,et al.  Multiplex PCR: Optimization and Application in Diagnostic Virology , 2000, Clinical Microbiology Reviews.

[7]  R. Allen,et al.  Polymerase chain reaction amplification products separated on rehydratable polyacrylamide gels and stained with silver. , 1989, BioTechniques.

[8]  V. Romanowski,et al.  Phenol extraction revisited: a rapid method for the isolation and preservation of human genomic DNA from whole blood. , 1994, Molecular and cellular probes.

[9]  E. Aberer,et al.  Importance of Sample Preparation for Molecular Diagnosis of Lyme Borreliosis from Urine , 2002, Journal of Clinical Microbiology.

[10]  Feng Xu,et al.  Analysis of DNA polymorphisms on the human Y‐chromosome by microchip electrophoresis , 2002, Electrophoresis.

[11]  R. Myers,et al.  Fine structure genetic analysis of a beta-globin promoter. , 1986, Science.

[12]  M. Kaku,et al.  Detection of mutations in adenine phosphoribosyltransferase (APRT) deficiency using the LightCycler system , 2000, Journal of clinical laboratory analysis.

[13]  D. Dash,et al.  Spectrum of β‐thalassemia mutations and their association with allelic sequence polymorphisms at the β‐globin gene cluster in an eastern Indian population , 2002 .

[14]  Wilson,et al.  Rapid, high‐throughput extraction of bacterial genomic DNA from selective‐enrichment culture media , 2001, Letters in applied microbiology.

[15]  K. Yoshimoto,et al.  Multiple fluorescence-based PCR-SSCP analysis using internal fluorescent labeling of PCR products. , 1996, BioTechniques.

[16]  Brian N. Johnson,et al.  An integrated nanoliter DNA analysis device. , 1998, Science.

[17]  P. Bastien,et al.  Comparison of Various Sample Preparation Methods for PCR Diagnosis of Visceral Leishmaniasis Using Peripheral Blood , 2001, Journal of Clinical Microbiology.

[18]  M. Caggana,et al.  Rapid, efficient method for multiplex amplification from filter paper , 1998, Human mutation.

[19]  A. Fox,et al.  Genotyping single nucleotide polymorphisms using intact polymerase chain reaction products by electrospray quadrupole mass spectrometry. , 2001, Rapid communications in mass spectrometry : RCM.

[20]  K. Mullis,et al.  Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. , 1987, Methods in enzymology.

[21]  M. A. Northrup,et al.  Functional integration of PCR amplification and capillary electrophoresis in a microfabricated DNA analysis device. , 1996, Analytical chemistry.

[22]  C. L. Liu,et al.  A simple method for DNA extraction from marine bacteria that produce extracellular materials. , 2003, Journal of microbiological methods.

[23]  L J Kricka,et al.  PCR in a silicon microstructure. , 1994, Clinical chemistry.

[24]  D. Ehrlich,et al.  Two-color multiplexed analysis of eight short tandem repeat loci with an electrophoretic microdevice. , 1999, Analytical biochemistry.

[25]  K. Young,et al.  Plastic microchip electrophoresis for analysis of PCR products of hepatitis C virus. , 1999, Clinical chemistry.

[26]  K. Mullis,et al.  Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. , 1985, Science.

[27]  Population genetic studies on nine tetrameric short tandem repeat loci using fluorescence dye‐labeled primers and capillary electrophoresis in the Austrian population , 1999, Electrophoresis.

[28]  Lihua Zhang,et al.  Effect of polymer matrix and glycerol on rapid single‐strand conformation polymorphism analysis by capillary and microchip electrophoresis for detection of mutations in K‐ras gene , 2005, Electrophoresis.

[29]  E. Rossomando,et al.  Capillary electrophoresis: separation and quantitation of reverse transcriptase polymerase chain reaction products from polio virus. , 1994, Journal of chromatography. B, Biomedical applications.

[30]  E. Scolnick,et al.  ras-Related gene sequences identified and isolated from Saccharomyces cerevisiae , 1983, Nature.

[31]  Marc Madou,et al.  MEMS-based sample preparation for molecular diagnostics , 2002, Analytical and bioanalytical chemistry.

[32]  Jinbao Zhu Use of PCR in Library Screening , 2002 .

[33]  P. Righetti,et al.  Capillary electrophoresis of polymerase chain reaction‐amplified products in polymer networks: The case of Kennedy's disease , 1994, Electrophoresis.