Development of genome-wide digital quantification method for non-coding RNAs which has single-base identification and single-molecule level quantification

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K14425
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Genome biology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: SHIROGUCHI KATSUYUKI 国立研究開発法人理化学研究所, 生命システム研究センター, ユニットリーダー (00454059)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: シークエンシング / デジタル計測 / バーコード

Outline of Final Research Achievements

Our goal was to develop a new digital RNA sequencing which could be applied for non-polyA tailed RNA and had single base resolution. Then, we aim to make a standardized high throughput digital RNA sequencing for researcher all over the world.
In this study, we attached one of many different molecular barcodes to each target molecules stochastically, amplified these molecules, and sequenced them. By counting the number of different barcodes, we quantified the original (i.e., before amplification) number of target molecules. Since the sequenced target molecules that have the same barcode sequence are from the same original molecules, we identified errors, and finally, we determined target sequences with single base resolution. I think this method will be used for quantification of nucleic acid molecules in many measurements.

Free Research Field

生物物理、定量ジェノミクス