Rationale: Prepared a TEM grid to obtain an image of the bacteriophage in the lysate

Procedure:

• Aseptic zone was set up to prevent bacterial contamination
• Pipetted 100μL of lysate from the original 50mL conical vial into a micro-centrifuge tube
• Traveled to the TEM lab
• Placed a strip of ParaFilm onto/into a agar plate
• Pipetted 20μL of lysate (in a drop) onto the ParaFilm, as well as two separate drops of 20μL DI water
• Pipetted one drop of 20μL Uranyl Acetate onto the ParaFilm strip
• Using precision forceps, placed a copper grid shiny side down into the lysate drop and let sit for five minutes
• After the allotted time (timed with a phone), transferred the grid into the first DI water drop and allowed to sit for two and a half minutes
• After the allotted time, transferred the grid into the other DI water drop and allowed to sir for two and a half minutes
• After the allotted time, transferred the grid into the Uranyl Acetate drop and allowed to sit for one minute
• After the allotted time, immediately extracted the grid from the Uranyl Acetate and wicked away extra liquid with a paper towel
• The prepared grid was put into the TEM and then imaged

Observations:

Image of the phage with dimensions measured

First image of the bacteriophage

• The measurement of the head was .055μL
• The measurement of the tail was .180μL

Conclusion/Next Steps: After getting this image, will be moving towards DNA extraction and the procedures following it, such as nano-drop and PCR.