Difference between revisions of "Culturing S2 Cells"
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| + | ==Materials== | ||
| + | *'''S2 cells''' see [[Freezing and Thawing S2 Cells]] | ||
| + | *'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) - stocked downstairs | ||
| + | *'''Penicillin/streptomycin''' 50X mix (Invitrogen, cat. no. 15240062) - stocked downstairs | ||
| + | *'''Insect Cell Media'''. Filter together Media (500 mL) and Pen/Strep (10 mL) into a tissue culture bottle | ||
| + | |||
| + | ==Protocol== | ||
| + | *Typically cells are split every 3-4 days at a ratio of 1:5 and grown at 25C | ||
| + | *After 2-3 days the cells become slightly less adherent. | ||
| + | #To split, gently aspirate the media (some cells will have detached from the plate surface) | ||
| + | #Pipet a gentle stream of media over the surface of the plate (5 mL for normal passage) | ||
| + | #Add 10 mL to fresh 10 cm dishes | ||
| + | #Add 1 mL of detached cells to each new plate | ||
| + | |||
| + | ==Knockdown== | ||
| + | *[[Designing and Preparing dsRNA]] | ||
| + | *[[dsRNA Mediated Knockdown in S2 Cells]] | ||
| + | |||
==References== | ==References== | ||
| − | see | + | see PMID 11752672 and PMID 18388942 |
| + | [[Category: Cell Culture]] | ||
| + | [[Category: Drosophila]] | ||
Latest revision as of 13:23, 3 August 2009
Contents
Materials
- S2 cells see Freezing and Thawing S2 Cells
- Sf-900 II SFM medium (Invitrogen, cat. no. 10902) - stocked downstairs
- Penicillin/streptomycin 50X mix (Invitrogen, cat. no. 15240062) - stocked downstairs
- Insect Cell Media. Filter together Media (500 mL) and Pen/Strep (10 mL) into a tissue culture bottle
Protocol
- Typically cells are split every 3-4 days at a ratio of 1:5 and grown at 25C
- After 2-3 days the cells become slightly less adherent.
- To split, gently aspirate the media (some cells will have detached from the plate surface)
- Pipet a gentle stream of media over the surface of the plate (5 mL for normal passage)
- Add 10 mL to fresh 10 cm dishes
- Add 1 mL of detached cells to each new plate
Knockdown
References
see PMID 11752672 and PMID 18388942
