RNA Sequencing by Direct Tagmentation of RNA/DNA Hybrids

Lin Di,Yusi Fu,Yue Sun,Jie Li,Lu Liu,Jiacheng Yao,Guanbo Wang, Yalei Wu,Kaiqin Lao,Raymond W. Lee, Genhua Zheng,Jun Xu,Juntaek Oh,Dong Wang,X. Sunney Xie,Yanyi Huang,Jianbin Wang

Proceedings of the National Academy of Sciences of the United States of America（2020）SCI 1区

Beijing Advanced Innovation Center for Genomics (ICG)

Cited 94|Views168

Abstract

Transcriptome profiling by RNA sequencing (RNA-seq) has been widely used to characterize cellular status, but it relies on second-strand complementary DNA (cDNA) synthesis to generate initial material for library preparation. Here we use bacterial transposase Tn5, which has been increasingly used in various high-throughput DNA analyses, to construct RNA-seq libraries without second-strand synthesis. We show that Tn5 transposome can randomly bind RNA/DNA heteroduplexes and add sequencing adapters onto RNA directly after reverse transcription. This method, Sequencing HEteRo RNA-DNA-hYbrid (SHERRY), is versatile and scalable. SHERRY accepts a wide range of starting materials, from bulk RNA to single cells. SHERRY offers a greatly simplified protocol and produces results with higher reproducibility and GC uniformity compared with prevailing RNA-seq methods.

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Key words

single cell,RNA-seq,Tn5 transposase

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