single-cell genomics

The isolation of single-cells remains a challenging task in single-cell genomics. Current methods lack the evidence that only a single-cell has been isolated in the analysis vessel. It is important that the integrity of the cells is maintained prior to their lysis in order to preserve their DNA.

Furthermore, cells should undergo as little stress as possible prior to their lysis in order to preserve their RNA and its expression level. The single-cell printer deposits single-cells in a very gentle manner, guaranteeing high purity and high viability.

This provides optimal basis for downstream single-cell genomic analysis.

workflow

Single cells of the osteosarcoma cell line U2OS were printed in wells of a 384-well microtiter plate, preloaded with 1 μl PBS. Additional cells were printed in dry wells resulting in a total single-cell printing efficiency of 98%.

Whole genome amplification (WGA) was performed on the cells and comparable DNA yields were achieved for dry and PBS wells. The WGA DNA was evaluated by a multiplex PCR on repetitive LINE1 transposons, which revealed positive results in all WGA samples.

In addition, U2OS-specific mutations in SLC34A2 (c.1538G>T), and in TET2 (c.1394C>T) were detected in representative WGA samples of single cells printed in PBS.

whole genome amplification

mutational analysis by sequencing

"Courtesy of the University of Freiburg - Julian Riba et al., Single-cell printing for the genetic analysis of cancer cells, Single Cell GenomicsMeeting 2015 Utrecht"