To prepare your samplesTo prepare your cell sample, please follow the instructions below:
Equilibrate the LUTHOR scd Buffer buffer at RT for 10min before diluting the cell sample
Filter the cell sample with the cell strainer (at least 1ml) to prevent clogging of the LUTHOR scd dispensing tip
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. Evaluate the % of cell debris and cell aggregates to set-up the thresholds for cell counting during dispense.
NOTE: Cell viability should be at least 90% to ensure best results for 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 150K 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 single cell dispenser)
Place the sample on the hold tube rack.
IMPORTANT: Keep your cells on ice until dispensing!