Prepping TEM Grid for Sample (Gabe) and results of Spot Test for Sample (Gabe) 11/19/18
Research Question:
To find out how the presence of bacteriophages in the soil around red or white oak trees has a correlation with the health condition of oak trees.
Rationale:
Individual phage particles are too small to be seen with light microscopes, necessitating the use of transmission electron microscopy (TEM). The Grid is vital to the fixing sample and staining them.
Experimental Procedure for TEM grid preparation for Soil (Gabe):
- High Titer Lysate
- 400-mesh copper grid
- TEM forceps
- DI water
- Uranyl Acetate
- Gloves
- p20 micropipette
- Parafilm
- Microcentrifuge tube
- Allocated 15 ul lysate sample, two individual H2O drops to the parafilm.
- Placed the copper grid on the lysate sample for 5 min.
- Placed the copper grid to the H2O for 2.5 min.
- Placed the copper grid to the other H2O for 2.5 min.
- Allocated one drop of Uranyl Acetate to the parafilm, and placed the grid onto the Uranyl Acetate for 1 min.
- Flick off redundant liquids on the copper grid and store away.
Observations, Results & Data:
The control plate was contaminated. The spotted regions on the sample plate were all completely lysed and appear to be clear.
Interpretations & Conclusions:
The sample plate was lysed on the spotted regions. Since the most diluted sample 10^-8 was also completely lysed that means the sample lysate 10^0 collected is extremely high titer. Therefore was able to start the TEM preparation.
Next Step:
The TEM preparation will continue, and the exact titer of the lysate sample would be calculated and DNA extraction would start.