Creating Cell Pellets¶
Created by Ravishankar Madhu¶
Last updated: 2023-07-10
Description¶
This protocol describes how to generate cell pellets from a cell suspension. This is applicable to cell lines and WBCs collected isolated from Peripheral Blood or Bone Marrow.
Reagents and Materials¶
- 1.5mL Eppendorf Tube
- Cell Solution
Things to Note before starting¶
Sample Processing¶
The 1.5mL Eppendorf tubes should be labeled with size 12 labels from the Label Maker adjacent to the Gel Bench. Cell pellets can be stored in -20C for months. Record where in the -20C you stored the cell pellets to prevent them from being lost.
Estimated Timing¶
This is a very quick procedure that takes 10 minutes on average. While the spinning time is fixed at 5 minutes, the aspiration time after pelleting increases linearly with the number of samples you have to pellet.
Procedure¶
- Determine how many cells you want to pellet. Common numbers people use are 10e6 if you have an abundance of cells. If your cell count is lower, add as much as you can.
- Aliquot that many cells into a 1.5mL Eppendorf tube.
- Spin down cells at 8000 rpm for 5 minutes on a tabletop centrifuge.
- Aspirate off the supernatant. Be careful to not disrupt the pellet at the bottom.
- Store cell pellets in the -20C Freezer.
Next Steps¶
Once cells are in cell pellets, their DNA can be extracted with a variety of different kits. Specific genes from the extracted DNA can be amplified by PCR.
Appendix¶
Related Protocols¶
- RM019 - DNA Extraction - Qiagen
- RM020 - DNA Extraction - Epicentre
- RM001 - PB or BM Lyse for WBC Isolation
- RM002 - CD34+ Enrichment