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. 10-40x10</sup>6
  2. Incubate at 37° C for 20-30 min.
  3. Add 0.1 volume resuspended DNase Inactivation Reagent.
  4. Incubate 2 min at RT mixing every 30s.
  5. Centrifuge at 10,000xg for 1.5 min.
  6. Transfer solution to a new tube avoiding pelleted Inactivation Reagent.

Notes

  1. List troubleshooting tips here.
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  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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