| Project/Area Number |
06558103
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Molecular biology
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| Research Institution | Kobe University |
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Principal Investigator |
ISONO Katsumi Kobe Univ., Faculty of Science Professor, 理学部, 教授 (70011640)
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| Co-Investigator(Kenkyū-buntansha) |
SAKURAI Toshiyuki Hitachi Electronic Engineering, Engineer, 技術本部研究部, 技師
HONJOU Atsuko Kobe Univ., Faculty of Science, Assistant, 理学部, 助手 (30229249)
KITAKAWA Madoka Kobe Univ., Faculty of Science, Assistant, 理学部, 助手 (70169853)
馬場 知哉 神戸大学, 理学部, 助手 (00338196)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥14,500,000 (Direct Cost: ¥14,500,000)
Fiscal Year 1995: ¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 1994: ¥8,600,000 (Direct Cost: ¥8,600,000)
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| Keywords | Transposon / PCR / Magnetic beads / Nucleotide sequencing / Lambda phage / Base-caller / Control sequence database / 対照,塩基配列データベース |
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| Research Abstract |
This project was aimed at establishing a method for more accurate and reliable base-calling system for the determination of nucleotide sequences using a DNA sequencer from Hitachi, model SQ3000. For this purpose, we obtained a large amount of nucleotide sequence raw data according to the transposon-facilitated PCR-based sequencing system of Kasai et al. (1992) using magnetic beads. As the template for analysis, we chose lambda phage DNA because of its easiness for preparation. We analyzed the distribution of peak distances and its dependence on the local nucleotide sequences and compared the sequence data obtained between electrophoresis lanes. The data were organized as a database and used for the probability calculation of individual nucleotides in the base-calling at each position of the sequence data. It thus turned out that the modified base-caller was more accurate in determining the sequence than the previous one : the accuracy level was at 97-100 % up to 300 bases and at 94-98 % up to 400 bases. Above 400 bases, however, the accuracy level was not very much improved by the new base-caller and it was found that other factors such as the quality of samples and acrylamide gel affected more on the base-calling accuracy.
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