Team:CU Boulder/Measurement


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InterLab Study

Devices Measured:

Protocols:

Interlab Parts

  1. Resuspend DNA from Distribution Kit using iGEM instructions..

    BBa_J23101: 2015 Kit Plate 1, Well 20K. Size: 35 bp.

    BBa_J23106: 2015 Kit Plate 1, Well 22A. Size: 35 bp.

    BBa_J23117: 2015 Kit Plate 1, Well 22K. Size: 35bp.

    BBa_I13504: 2015 Kit Plate 4, Well 21J. Size: 875 bp.

  2. Transform BioBrick parts using Transformation Protocol.
  3. Confirm transformation
    1. Fill culture tubes with 5 mL of LB and appropriate amount and type of antibiotic.
    2. Pluck colonies from plates using 200 μL pipette tip and place in tubes, leave tubes in 37.5 ° shaking incubator on 250 rpm for 12-18 hours.
    3. Retrieve overnight cultures from incubator and miniprep using Qiagen Miniprep Protocol
    4. Digest miniprepped DNA using Spe1 and Pst1 restriction enzymes
    5. Run a gel and image to validate construct.
  4. Create construct.
    1. Digest BBa_J232101 with SpeI and PstI at 37 °C for 60 minutes.
    2. PCR I13504 with appropriate primers and digest using Xba1 and Pst1.
    3. Gel extract digested DNA.
    4. Ligate both pieces of DNA using gel image and ligation protocol
    5. Repeat with BBa_J23106 and BBa_J23117
  5. Transform ligation product.
  6. Confirm transformation
    1. Fill culture tubes with 5 mL of LB and appropriate amount and type of antibiotic.
    2. Pluck colonies from plates using 200 μL pipette tip and place in tubes, leave tubes in 37.5 ° shaking incubator on 250 rpm for 12-18 hours.
    3. Retrieve overnight cultures from incubator and miniprep using Qiagen Miniprep Protocol
    4. Digest miniprepped DNA using Spe1 and Pst1 restriction enzymes
    5. Run a gel and image to validate construct.

Data:

J23101

  • Colony 1
    1. Mean = 3243.76
  • Colony 2
    1. Mean = 2794.08
  • Colony 3
    1. Mean = 5838.96

    J23106

  • Colony 1
    1. Mean = 10.53
  • Colony 2
    1. Mean = 6.68
  • Colony 3
    1. Mean = 6.70

    J23117

  • Colony 1
    1. Mean = 4.72
  • Colony 2
    1. Mean = 3.83
  • Colony 3
    1. Mean = 3.89
    • Team:CU-Boulder - 2015.igem.org