Direct Cell Lysis (plate-based) Single Cell Sequencing

Single Cell Full Length RNA Sequencing

For full transcriptome single-cell sequencing, the AGC offers plexWell Rapid Single Cell (seqWell) processing. Users will sort cells into individual wells containing lysis buffer on a 96-well plate. After sorting, the plates can be frozen until such time that the user is ready to submit samples to the core for library prep and sequencing. Library prep uses a template-switching method to generate full-length cDNAs (SMART-seq) and subsequent library construction. This process provides full-length transcript coverage, allowing for more in-depth analyses, including variant calling and isoform detection.

Image provided by seqWell

Assay Requirements

•       Kit specific lysis buffer (pre-aliquoted plates can be picked up from the AGC)
•       Sort cells in ≤5 µl of nuclease-free water or ≤1 µl of nuclease-free water PBS

Cells should be deposited directly into the lysis plate. We strongly encourage the incorporation of a positive and negative control. The negative control should be an aliquot of your cell sorting buffer. The positive control well (H12) should be left blank for AGC staff to place a control RNA sample.

The plate (with deposited cells) is stable at -80 for up to three months so can safely be transferred back to the core on dry ice.