Analysis of radiation induced mutations using next generation sequencer

2014 Fiscal Year Final Research Report

• Project/Area Number: 25550030
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Hiroshima University
• Principal Investigator: KANAI Akinori 広島大学, 原爆放射線医科学研究所, 助教 (60549567)
• Co-Investigator(Kenkyū-buntansha): KAWAI Hidehiko 広島大学, 原爆放射線医科学研究所, 助教 (30379846)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 放射線 / 次世代シーケンサー

Outline of Final Research Achievements

To estimate radiation induced mutation frequency with single-cell sequencing, HT1080 single cells were sorted and gamma-irradiated 0.4 Gy and 2 Gy. The cell divided several times, DNA was extracted from each single cell and we performed whole genome amplification (MALBAC method), exon capture and NGS sequence.
The number of 2 Gy specific base substitutions was 20 and indels were 3 in 7.5 Mbp. The number of 0.4 Gy specific base substitutions was 27 and indels were not detected in 14 Mbp.

Free Research Field

分子生物学