| Natalie Raso - Weapons of Targeted Destruction: Using Viruses to Kill Cancer | Procedure for KM110r Infection |
| Project
Information Abstract Project Summary Background Purpose Scientific Thought Hypotheses Apparatus and Materials Genetically Engineered KM110red Herpesvirus Methodology Procedure for Cell-Line Splitting Procedure for KM110r Infection Procedure for Immunofluorescent Microscopy Imaging Statistical Analyses Proliferation Assay Analyzed Data Major Results Graphed Results Discussion of Statistics Controls and Variables Conclusions Discussion Discussion of KM110r Efficacy Successes and Failures Sources of Error and Data Limitations Future Research Applications Glossary Bibliography Acknowledgements |
24 hours after the cells were split
to a 6-well dish, they were prepared for infection with KM110r. Since
there were two wells each of hFOB, U2OS and MIX, one of each was infected,
and the other was mock-infected as the negative control. This procedure
was carried out in a Level II Biohazard Tissue Culture Laboratory in a
Level II Biohazard Cell Hood. The steps for performing this procedure are
outlined below. Initial
Calculations Inoculumà 400 µL
SF-DMEM KM110rà 3 x 107 particle forming units (PFU) per
mL Multiplicity of
Infection (MOI)à
1 Volume of KM11or = 1.5 x 105
cells x 1
PFU per/cell
3x107 PFU /
mL
= 0.41 x 10-2 mL
= 0.41 x 10-2 1000µL
= 4
µL 1.
Remove
media from each well with aspirating
pipette. 2.
Add
400µl of SF-DMEM to all 6
wells. 3.
Add 4µL thawed stock of
KM110r to right (infected side only). 4.
Incubate at desired temperature
for 5 x 6 minute intervals, rocking the cells after each
interval. 5.
Remove virus using aspirating
pipette. 6.
Add 2mL DMEM + 10% FBS to all 6
wells. |