title: A highly sensitive and accurate gene expression analysis by sequencing ("bead-seq") for a single cell.
abstract: 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.
authors.affiliation: Hitachi Central Research Laboratory, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Matsunaga
authors.affiliation: Hitachi Central Research Laboratory, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Goto
authors.affiliation: Hitachi Central Research Laboratory, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Arikawa
authors.affiliation: Hitachi Central Research Laboratory, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Shirai
authors.affiliation: Hitachi Central Research Laboratory, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsunoda
authors.affiliation: First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Huang
authors.affiliation: Hitachi Central Research Laboratory, Kokubunji-shi, Tokyo 185-8601, Japan. Electronic address: hideki.kambara.se@hitachi.com.
authors.forename: Hideki
authors.initials: H
authors.lastname: Kambara
mesh_terms: D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D045325:HCT116 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA / Q000379:methods*; D059010:Single-Cell Analysis
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger
keywords: Gene expression; Single cell; mRNA-seq