single-cell genomics

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

The typical work flow in single-cell genomics applications contains four major steps. From the cell to the genomic information. cytena is expert to help you with the challenges in step 1 - the isolation of individual cells from your sample.

isolation

individual cells of interest have to be isolated efficiently and automated

lysis

The isolated cell has to lysed to extract their DNA, RNA, etc.

amplification

The lysate has to amplified to gain sufficient material to start the analysis

sequencing

the amplified material is sequenced to read the genetic information

application example

Single cells of the osteosarcoma cell line U2OS were dispensed in wells of a 384-well microtiter plate, preloaded with 1 μl PBS. Additional cells were dispensed in dry wells resulting in a total single-cell dispensing 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 dispensed 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"