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)
Dilute the cells to the recommended concentration (see FAQ: What are the cell sample concentration requirements?)
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 step is required before staining.
Dilute the sample ideally to 150,000 cells/ml
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!