GHV-68 Plaque Assay

Overview

Plaque assay for gHV-68 using 3T12 fibroblasts. Stolen from Virgin lab protocols.

Materials

• Methylcellulose
• X μL reagent 2
  • component A (reagent 2 is made up of multiple components)
  • component B
• equipment 1
• equipment 2

Procedure

Day 0

1. Trypsinize 3T12's (1 confluent T175 will yield approx. 10-40x10^6, cells) and resuspend at 2.5-2.5x10^{6<\sup>/ml in DMEM-10.}
2. 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.
   1. Need one 6-well plate per viral sample. (This will let you test 6 logs of dilution)
3. PrepareMethylcellulose, if necessary.
4. Freeze/thaw/freeze your viral sample, if necessary.

Notes

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References

Relevant papers and books

1. Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. DOI:10.1073/pnas.78.11.6840 |
2. JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. DOI:10.1016/s0022-2836(61)80072-7 |
3. ISBN:0879697164

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