A highly sensitive and accurate gene expression analysis by sequencing ("bead-seq") for a single cell

作者：Matsunaga Hiroko; Goto Mari; Arikawa Koji; Shirai Masataka; Tsunoda Hiroyuki; Huang Huan; Kambara Hideki^*

来源：Analytical Biochemistry, 2015, 471: 9-16.

DOI：10.1016/j.ab.2014.10.011

• Single cell
• Gene expression
• mRNA-seq

Analyses of gene expressions in single cells are important for understanding detailed biological phenomena. Here, a highly sensitive and accurate method by sequencing (called "bead-seq") to obtain a whole gene expression profile for a single cell is proposed. A key feature of the method is to use a complementary DNA (cDNA) library on magnetic beads, which enables adding washing steps to remove residual reagents in a sample preparation process. By adding the washing steps, the next steps can be carried out under the optimal conditions without losing cDNAs. Error sources were carefully evaluated to conclude that the first several steps were the key steps. It is demonstrated that bead-seq is superior to the conventional methods for single-cell gene expression analyses in terms of reproducibility, quantitative accuracy, and biases caused during sample preparation and sequencing processes.

• 出版日期2015-2-15
• 单位南京医科大学