| Project/Area Number |
14370071
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Human pathology
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| Research Institution | Yamaguchi University |
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Principal Investigator |
SASAKI Kohsuke YAMAGUCHI UNIV., SCH.OF MED PATHOLOGY, Professor, 医学部, 教授 (80116722)
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| Co-Investigator(Kenkyū-buntansha) |
OGA Atunori YAMAGUCHI UNIV., SCH.OF MED PATHOLOGY, RESEARCH ASSOCIATE, 医学部, 助手 (90243633)
KAWAUCHI Shigeto YAMAGUCHI UNIV., SCH.OF MED PATHOLOGY, ASSISTANT PROFESSOR, 医学部, 講師 (80284511)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 2003: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 2002: ¥7,200,000 (Direct Cost: ¥7,200,000)
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| Keywords | DNA / SNP / Flow cytometry (FCM) / Hybridization / fluorescence / high-throughput analysis / drug response / genotype / フローサイトメトリー / ゲノム / フローアレイ / 蛍光 / SNPs |
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| Research Abstract |
DNA sequence variation is associated with a certain disease and differential drug response. Single nucleotide polymorphisms (SNPs) are common type of variant. In this project, we have developed a new method for high-throughput genotyping of SNPs with microspheres of various sizes and a flow cytometer equipped with only a single laser
This system uses microspheres of various sizes with oligonucleotide probes attached by covalent bonds. Polymerase chain reaction (PCR) products labeled with Alexa 488 are hybridized to the probes and then hybridization signals are analyzed by flow cytometry. In order to type a specific SNP, two kinds of beads are bound to paired probes ; one complementary to the wild-type sequence and the other to the variant sequence. The paired beads are placed in the same tube and the number of normal beads and variant beads ratio are three to one. When the beads with higher fluorescence intensity are about three times as many as lower ones, this sample is determined wild-type genotype. When multiplex analysis is carried out, the beads for each SNP site is identified by their diameters. In this project, we have developed a novel method in which SNP genotyping with microspheres of various sizes and flowcytometer equipped with single laser. The method increases parameters in beads analysis
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