How should I prepare my cell sample?

To prepare your cell sample, please follow the instructions below:

• Equilibrate the LUTHOR scd buffer at RT for 10min before diluting the cell sample

• Filter the cell sample with the cell strainer (at least 1ml)

• Check cell concentration and viability using a cell counter (with Trypan Blue staining, Dual Acridine Orange (AO) staining or Propidium Iodide (PI) staining), fluorescent cell counter or microscope.

NOTE: Cell viability should be at least 90% to ensure best results with the LUTHOR HD library preparation.

IMPORTANT: If you are working with fixed cells, cell viability should be determined before fixation! Additionally, fixed, non-permeabilized, dead cells will not all be stained with Propidium Iodide. In order to count fixed cells, permeabilization is required before staining!

• Dilute the sample ideally to the recommended concentration 150,000 cells/ml (see FAQ: What are the cell sample concentration requirements?)

• Add the LUTHOR scd buffer to the sample and pipet slowly for mixing (by using the wide-bore filter tips provided with the LUTHOR single cell dispenser)

• Place the sample on the hold tube rack.

IMPORTANT: Keep your cells on ice until dispensing!