Optimization of whole genome amplification methods for single cell analysis
| Project/Area Number |
16K11106
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Keio University |
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Principal Investigator |
SATO Suguru 慶應義塾大学, 医学部(信濃町), 助教 (40383898)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 着床前遺伝子診断 / シングルセルPCR / 全ゲノム増幅法 / アレルドロップアウト現象 / 全ゲノム増幅技術 / 単一細胞解析 / single cell PCR / 着床前診断 / 染色体マイクロアレイ解析 |
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| Outline of Final Research Achievements |
It is known that the allele dropout phenomenon, in which only one of the alleles is not amplified, occurs in the process of genome amplification from scarce amount of DNA and causes a misdiagnosis. We compared various whole genome amplification techniques (WGA) available. We observed that the mutations detected differed depending on the each WGA methods. Amplification bias was observed even in the same gene region. ADO was observed at any locus regardless of which WGA was adopted. At present, the MDA method is the most versatile.
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| Academic Significance and Societal Importance of the Research Achievements |
単一遺伝子病のための着床前遺伝子診断は, 今尚国内における実施数は多くないが, 次世代シーケンサをはじめとする遺伝子工学技術やタンデムマススクリニーング事業の導入により, 今後今まで原因不明とされた稀少遺伝子疾患や代謝異常症のための着床前診断の要望が増えてくると思われる. 全ゲノム増幅法の導入により, より簡便にシングルセルPCRを実施できる効果が期待される. 本研究では, 家系毎に異なる遺伝子異常の変異に応じた全ゲノム増幅技術の選択の最適化のために, 有用な情報を提供するものと考えられる.
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Report
(5 results)
Research Products
(5 results)
