Sample Preparation
Sample Submission
Suspension cells
Collect cells by centrifugation for 5 minutes at 1100 rpm, remove supernatant and dilute cells to a concentration of 1x106 cells/ml in fresh pre-warmed medium. Immediately transfer your cells to the GARP Core (with a scheduled appointment).
Adherent cells
Adherent cells can be removed by enzymatic digestion using 1 ml of pre-warmed 0.25% or 0.05% trypsin. Trypsinize for 1 minute or longer at RT depending on your cell type to ensure that the cells are unclumped and surface glycoproteins remain intact.
After cell detachment, add 4 ml of fresh pre-warmed medium to 1 ml of trypsinized cells and centrifuge for 5 minutes at 1100 rpm. Remove supernatant and re-suspend cells in fresh pre-warmed medium.
Dilute cells to a concentration of 1x106 cells/ml in fresh medium. Immediately transfer your cells to the GARP Core (with a scheduled appointment).
Cryopreservation cells
Cells should be frozen under conditions that minimize lysis. Supplement your cell culture media with 10% DMSO and dilute cells to a concentration of 1x106 cells/ml, put 1 ml of the cell suspension into a cryovial and slow-freeze (-1oC per minute) in an isopropanol-filled container placed at -80oC, using a device such as 'Mr. Frosty' or an equivalent.
Note: Depending on your cell type(s), cryopreservation conditions may need to be optimized to ensure that cell viability remains greater than 95% after one week stored at -80oC.
Tissues
Contact GARP core at garpcore@bcmm.edu for more information.
Cell Submission
- Cell viability must be > 95%
- When providing cells for your CUT&RUN or ATAC-seq projects, include an additional 30% for quality control checks.
- Live cells should be delivered in fresh medium, kept on ice with a scheduled appointment.
- Cryopreserved cells must be transported on dry ice and dropped off the cells in R.325E.
