Team:BostonU/Notebook

Our Project
Day by Day Protocols

Notebook

May

Week One - May 28th to May 29th
  • May 28th 2015
    1. Made broth and agar
    2. Plated the agar
    3. Aliquot 50mL LB media tubes
    4. Innoculate Ben’s mammalian backbones with dimerization domains and Ben’s phiC-31, TP901-1, ofr7 and gp3 plasmids
  • May 29th 2015
    1. Cell stock creation from frozen plasmids
    2. Miniprepped the plasmids we grew up and cell stocked for iGEM cell stocks
    3. Using the nanodrop fluorospectrometer
Week Two - June 1st to June 5th
  • June 1st 2015
    1. Digested Ben's backbones
    2. PCR set-up
    3. Made an agarose gel to separate the products of the PCR
    4. Made more pS-5 and pS-18 cell stocks (Our pS-5 and pS-18 cell stocks gave us a very low yield when their concentrations were tested with a nanodropper).
  • June 2nd 2015
    1. Mini-prepped the pS-5 and pS-18 cell stocks that were made yesterday.
    2. Measured the concentrations of DNA from the purified digestion results and purified gel results.
    3. Digested pS-5 because it is a backbone. pS-18 is not a backbone that is why it was not re-digested.
    4. Ran the PCR for pS-14 and 10 different primers for a second time, the first time we did this was yesterday but the test yielded strange results in that our bands were thick, clouded, and our ladder was relatively useless because it did not run for a long enough period of time.
    5. Ran the gel for the results from today’s PCR in order to see if we can get better results than yesterday and we did.
    6. Gel purified
    7. Digesting Our Two Integrases with Enzymes in Order to Ligate (the pS-14 phi 31 integrases into pS-1:12 backbones)
    8. PCR Purification of Our Restriction Results to Get the Integrase Inserts
    9. Ligation
  • June 3rd 2015
    1. Checked out the colonies and Ben found that our pS-9 and pS-11 had very low yields when checked against their respective negative control colonies
    2. Picked colonies
    3. Emailed former BU students who developed a light stimulation device
  • June 4th 2015
    1. Took colonies out of incubator, decanted media
    2. Miniprepped from deep wells
    3. Test cut plasmids
    4. Ran gel electrophoresis
Week Three - June 8th to June 12th
  • June 8th 2015
    1. Repicked colonies for bad test cuts
      1. 200, 201, 204, 205, and 208
      2. Created new deep well plates with LB + Carb
      3. Picked three new colonies from each of these plates
      4. Incubated reference plate and deep well plate overnight
    2. Saving a Gel (There was an incorrect number of wells in a gel we made so: )
      1. We tore up the gel and put it into the iGEM gel erlemyer flask
      2. Microwaved the 100mL of gel for one minute until it was melted
      3. We re-added the ethidium bromide
      4. We re-poured the gel
  • June 9th 2015