Team:Stony Brook/Notebook/Vaccine-W2

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Week 2: 7/5-7/8

Week 2 (7/5-7/8)



7/5

  • Things

7/6

Ran a digest on two plasmids
  • Both were incubated for 2 hours at 37°C
Reaction A
    Content ul
    Plasmid 4.55
    PstI 1
    BsmbI 1
    3.1 pH Buffer 5
    Nuclease-free water 38.45
Reaction B
    Content ul
    Plasmid 4.55
    PstI 1
    3.1 pH Buffer 5
    Nuclease-free water 39.45
Ran gel of PCR'ed Gal and Construct from IDT

Lanes:

  1. Ladder
  2. Gal (Brian
  3. Construct (Brian)
  4. Gal (Noah)
  5. Construct (Noah)
  • Ran at 100V for 1 hour
Second gel of digests A and B from above

Lanes:

  1. Ladder
  2. A
  3. B
  4. A
  5. B
  • Used 15ul of each reagent
  • Ran at 100V for 1 hour
Nanodraopped gel extraction for the PCR'ed Gal and Construct
    Content Concentration (ug/ul)
    Gal (Brian) - E 28.7
    Construct (Brian) - G 7.2
    Gal (Noah) - F 64.2
    Construct (Noah) - H 11.3
PCR Gal and Construct together
    Contents ul
    Phire 25
    Fwd Primer 2.5
    Rev Primer 2.5
    Gal1 - N 0.389
    Construct - N 2.21
    Nuclease-free Water 17.40
PCR Gal and Construct together
    Contents ul
    Phire 25
    Fwd Primer 2.5
    Rev Primer 2.5
    Gal1 - B 0.87
    Construct - B 3.47
    Nuclease-free Water 15.66

7/7

  • Poured 2 culture tubes with ampicillin and ASK BRIAN ABOUT THAT
Gel of PCR'ed combined Gal1 and Construct

Lane:

  1. Ladder
  2. Empty
  3. Brian
  4. Noah
  • Ran at 100V for 1 hour
Ran a PCR

Program settings:

    Phase Temperature (°C) Time (s)
    Initial Denaturation 98 30
    Denaturation 98 20
    Annealing 63 20
    Extension 72 60
    Final Extension 72 60
    • Ran a gel on the product