GHV-68 Plaque Assay
Plaque assay for gHV-68 using 3T12 fibroblasts. Stolen from Virgin lab protocols.
- X μL reagent 2
- component A (reagent 2 is made up of multiple components)
- component B
- equipment 1
- equipment 2
- Trypsinize 3T12's (1 confluent T175 will yield approx. 10-40x106, cells) and resuspend at 2.5-2.5x106/ml in DMEM-10.
- Aliquot 2/ml/well (3-5x105 cells/well) into 6-well plates. Don't swirl the cells, as you may end up with all your 3T12's in the center of the well.
- Need one 6-well plate per viral sample. (This will let you test 6 logs of dilution)
- Prepare Methylcellulose, if necessary.
- Freeze/thaw/freeze your viral sample, if necessary.
- List troubleshooting tips here.
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- Anecdotal observations that might be of use to others can also be posted here.
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Relevant papers and books
- Goldbeter A and Koshland DE Jr. . pmid:6947258.
- JACOB F and MONOD J. . pmid:13718526.
- Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164.
- Who has experience with this protocol?
or instead, discuss this protocol.