October
26
OCTOBER 22ND AND 24TH- Labs
- OCTOBER 22ND, 2018
- OBJECTIVE:
- Conduct a gel electrophoresis to test if the PCR was positive
- PROCEDURE:
- .8 grams of the agarose powdered was measured out
- Then 40mL of 1X TAE was put into a flask
- The agarose powder was then poured into the flask, the flask was then repeatedly swirled
- The flask when then placed in the microwave, and heated until it started to boil
- The flask was then taken out and swirled until it cooled, and was just warm (instead of burning hot)
- Then 2𝝁L EtBr was added by a Teaching Assistant
- The solution was then poured into a gel apparatus, where a gel comb was then inserted
- Then 20 minutes was allowed for gel to cool
- Gel comb was removed
- Then the box holding the gel was filled with 1X TBE to submerge to gel
- Next 10 𝝁L of each sample was loaded into the appropriate wells seen in the figure below:
- The lid was then placed on the box, and the power cords were connected to the power apparatus, and set at 100 volts (make sure positive end is on end away from wells)
- This was run for 40 minutes, then gels were taken to MCB to be imaged by Teaching Assistant
- Imaging was done by cleaning the UV plate the with ethanol, placing the gel on it, then placing it in the BioRad Gel Doc EZ imaging machine
- RESULTS:
- The control gel results came out normal, or as expected
- The plate ran with all the samples was negative for any DNA presence
- CONCLUSION:
- Since there was no DNA present in the gels, it can be determined that the soil samples obtained do not contain phage
- NEXT STEPS:
- Collect new sample, then wash sample to run another PCR
- OBJECTIVE:
- OCTOBER 24TH, 2018
- OBJECTIVE:
- Wash new soil sample to prepare to run PCR
- PROCEDURE:
- Soil was filled to the 2mL mark of a test tube
- Then 10mL of LB broth was added to the tube
- It was shaken fro 15 minutes, before then bing placed into the centrifuge for 10 minutes where it was then spun at 3,000G
- Next a well plate was obtained, and 2mL of the supernatant was placed into each of the three wells
- The well plate was then heated to 55C for 5 minutes
- The sample was then cooled or 5 minutes
- Then .5 𝝁L of arthro was added to each well
- The well plate was then incubated at 28C and were wired at 150 RPM
- RESULTS:
- No Results to report
- CONCLUSION:
- No Results to report
- NEXT STEPS:
- Run PCR
- OBJECTIVE:
