Development of a reference material to quantify only a few DNA molecules

• Project/Area Number: 26450185
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Food science
• Research Institution: National Agriculture and Food Research Organization
• Principal Investigator: TAKABATAKE Reona 国立研究開発法人農業・食品産業技術総合研究機構, 食品研究部門 食品分析研究領域, 上級研究員 (20463466)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 定量分析 / PCR / DNA / 分子 / 遺伝子組換え

Outline of Final Research Achievements

In this study, I attempted to develop a reference material containing only a few copy number of PCR target sequences to evaluate existing DNA quantification techniques. To develop the reference material, a standard DNA, designated as standard DNA-N, in which PCR-targeted sequences were repeatedly connected in tandem, was prepared. In the standard DNA-N, recognition sequences for several restriction endonuclease were located between of each PCR-targeted sequence. The DNA solution was highly diluted to make the average number of the standard DNA-N molecules in a well below one, subsequently, the diluted solutions were treated with restriction endonucleases, and then the reference material just containing N copies of the PCR-target sequences could be obtained. I have already prepared the standard DNA-16.