GHV-68 Plaque Assay

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Contents

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. 10-40x106,<\sup> cells) and resuspend at 2.5-2.5x106<\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

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

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References

Relevant papers and books

  1. Goldbeter A and Koshland DE Jr. . pmid:6947258. PubMed HubMed [Goldbeter-PNAS-1981]
  2. JACOB F and MONOD J. . pmid:13718526. PubMed HubMed [Jacob-JMB-1961]
  3. Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164. [Ptashne-Genetic-Switch]
All Medline abstracts: PubMed HubMed

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.

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