Marek: Freeze-down/Thaw: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 7: | Line 7: | ||
*10-50 ml centrifuge tube | *10-50 ml centrifuge tube | ||
*DMSO, Tissue culture (TC)-tested (e.g. Sigma, D2650) | *DMSO, Tissue culture (TC)-tested (e.g. [http://www.sigmaaldrich.com Sigma], D2650) | ||
*Cell culture medium (depends on cell line/cells) | *Cell culture medium (depends on cell line/cells) | ||
*Trypsin-EDTA (e.g. from [http://www.paa.com PAA], L11-004) - for adeherent cells | *Trypsin-EDTA (e.g. from [http://www.paa.com PAA], L11-004) - for adeherent cells |
Revision as of 02:54, 18 May 2007
Overview
This short protocol describes how to freeze down and thaw (or bring up) mammalian cells (e.g. HeLa, KEK293, CHO, Jurkat T cells).
Materials
List reagents, supplies and equipment necessary to perform the protocol here.
- 10-50 ml centrifuge tube
- DMSO, Tissue culture (TC)-tested (e.g. Sigma, D2650)
- Cell culture medium (depends on cell line/cells)
- Trypsin-EDTA (e.g. from PAA, L11-004) - for adeherent cells
- Freeze-down medium
- 6% DMSO
- Cell culture medium (you can use 100% fetal calf serum (FCS) instead of cell culture medium)
- Basic TC centriguge (0 - 2500 rpm)
- Freeze-down box (this could be simply made from a small styropore box and cotton or from two styropore racks from 15-ml centrifuge tubes )
Procedure
- Step 1
- Step 2
- Step 2 has some additional information that goes with it. i.e. Keep at 4°C.
- Step 3
- Step 3 has multiple sub-steps within it.
- Enumerate each of those.
Notes
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.