October 26

Electrophoresis on Sample Enrichment Lysate (10.1.18) after PCR 10/21/18

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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:

By implementing Electrophoresis, the image captured on the gel after could help determine if there is any trace of DNA amplified after PCR in the filtered Enrichment Lysate sample.

Electrophoresis for sample (10.1.18):

Materials:

  • 1x TBE
  • Agarose powder
  • EtBr
  • Electrophoresis apparatus and power supply
  • Heat source
  • DNA loading dye
  • DNA ladder
  • Gel Imager

Procedure:

  1. prepare a solution with 40 ml of 1x TBE and 0.8 g Agarose
  2. microwave until the solution boils and retrieves it for cooling
  3. Add EtBr to the solution and have a final concentration of 0.5 %
  4. While the solution is still warm, pour it into the preassembled gel apparatus
  5. wait for solidification
  6. After the gel had solidified, fill the apparatus with 1x TBE
  7. Load the samples(10 ul), DNA dye(10 ul) and DNA ladder(5 ul) into the wells
  8. Run the gel at 100 volts 45 min

Observations, Results & Data:

The result from imaging shows no significant bands anywhere on the gel, indicating there is no significant amount of DNA in the filtered enrichment sample lysate after PCR.

Interpretations & Conclusions:

The lack of distinct bands on the image shows that there are no phage DNA present in the lysate sample, however, it is notable that the bottom part of the gel has traces of band, it might be the primers replicated themselves during PCR.

Next Step:

Due to the negative results of the PCR Electrophoresis experiment, the new objective id to collect more samples to continue the search for phages.


Posted October 26, 2018 by joseph_yu1 in category Yang-En Yu

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