G&T-Seq

G&T-Seq

G&T-Seq can separate and sequence genomic DNA and full-length mRNA from single cells. In this method, single cells are isolated and lysed. RNA is captured using biotinylated oligo(dT) capture primers and separated from DNA using streptavidin-coated magnetic beads. Smart-seq2 is used to amplify captured RNA on the bead, while multiple displacement amplification (MDA) is used to amplify DNA. After sequencing, integrating the DNA and RNA sequences provides insights into the gene expression profiles of single cells.

Pros:

• Compatible with any whole-genome amplification method
• No 3'-end bias in sequence reads because full-length transcripts are captured
• Because DNA and RNA are physically separated and amplified independently, there is no need to mask coding sequences during analysis

Cons:

• Physical separation of DNA and RNA can increase the risk of sample loss or contamination
• Physical separation of DNA and RNA increases handling time

• G&T-Seq: Macaulay I. C., Haerty W., Kumar P., Li Y. I., Hu T. X., et al. (2015) G&T-seq: parallel sequencing of single-cell genomes and transcriptomes. Nat Methods 12: 519-522

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