Dissolvable Polyacrylamide Beads for High-Throughput Droplet DNA Barcoding

被引：31

作者：

Wang, Yongcheng ^{[1
,2
,3
,4
]}

Cao, Ting ^{[1
,2
,3
,5
]}

Ko, Jina ^{[1
,6
]}

Shen, Yinan ^{[2
,3
]}

Zong, Will ^{[2
,3
]}

Sheng, Kuanwei ^{[7
]}

Cao, Wenjian ^{[7
]}

Sun, Sijie ^{[2
,3
]}

Cai, Liheng ^{[2
,3
]}

Zhou, Ying-Lin ^{[5
]}

Zhang, Xin-Xiang ^{[5
]}

Zong, Chenghang ^{[7
]}

Weissleder, Ralph ^{[6
,8
]}

Weitz, David ^{[1
,2
,3
]}

机构：

[1] Harvard Univ, Wyss Inst Biol Inspired Engn, Boston, MA 02115 USA

[2] Harvard Univ, John A Paulson Sch Engn & Appl Sci, Cambridge, MA 02138 USA

[3] Harvard Univ, Dept Phys, Cambridge, MA 02138 USA

[4] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA

[5] Peking Univ, BNLMS, MOE Key Lab Bioorgan Chem & Mol Engn, Coll Chem & Mol Engn, Beijing 100871, Peoples R China

[6] Harvard Med Sch, Massachusetts Gen Hosp, Ctr Syst Biol, Boston, MA 02114 USA

[7] Baylor Coll Med, Dept Mol & Human Genet, Houston, TX 77030 USA

[8] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA

来源：

ADVANCED SCIENCE | 2020年 / 7卷 / 08期

关键词：

barcode beads; dissolvable hydrogels; droplet microfluidics; single-cell sequencing; SINGLE-CELL TRANSCRIPTOMICS; RNA-SEQ;

D O I：

10.1002/advs.201903463

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Droplet-based single cell sequencing technologies, such as inDrop, Drop-seq, and 10X Genomics, are catalyzing a revolution in the understanding of biology. Barcoding beads are key components for these technologies. What is limiting today are barcoding beads that are easy to fabricate, can efficiently deliver primers into drops, and thus achieve high detection efficiency. Here, this work reports an approach to fabricate dissolvable polyacrylamide beads, by crosslinking acrylamide with disulfide bridges that can be cleaved with dithiothreitol. The beads can be rapidly dissolved in drops and release DNA barcode primers. The dissolvable beads are easy to synthesize, and the primer cost for the beads is significantly lower than that for the previous barcoding beads. Furthermore, the dissolvable beads can be loaded into drops with >95% loading efficiency of a single bead per drop and the dissolution of beads does not influence reverse transcription or the polymerase chain reaction (PCR) in drops. Based on this approach, the dissolvable beads are used for single cell RNA and protein analysis.

引用

页数：6

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