Team:Lethbridge Canada/notebook march

From 2013hs.igem.org

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<li class="notebook_link_title"><a href="notebook_june.html">June</a></li>
<li class="notebook_link_title"><a href="notebook_june.html">June</a></li>
<li><a href="notebook_june.html#entry_one">June 1, 2013: Restrictions</a></li>
<li><a href="notebook_june.html#entry_one">June 1, 2013: Restrictions</a></li>
<li><a href="notebook_june.html#entry_two">June 2, 2013: Transformation</a></li>
<li><a href="notebook_june.html#entry_two">June 2, 2013: Transformation</a></li>
<li><a href="notebook_june.html#entry_three">June 4, 2013: Picked Colonies</a></li>
<li><a href="notebook_june.html#entry_three">June 4, 2013: Picked Colonies</a></li>
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<li><a href="notebook_june.html#entry_four">June 10, 2013: Sent constructs for sequencing</a></li>
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<li><a href="notebook_june.html#entry_five">Transformation of OXY into BL21</a></li>
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<li><a href="notebook_june.html#entry_six">Restricted and Ligated and Transformed</a></li>
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<li><a href="notebook_june.html#entry_seven">Picked Cells</a></li>
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Revision as of 03:15, 16 June 2013

Transformations

  • Name: Marie C, Fiona S, Chris I, Katie T, Wesley M
  • Date: March 12, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Antibiotic: Kan and Amp
  • Part Number and Plasmid: R0010 PSB1A2, B0032 PSB1A2, J04650 PSB1AK3, B0031 PSB1A2, B0030 PSB1A2, J04500 PSB1AK3
  • Results: TBA

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Picking Cell Colonies (Cultures)

  • Names: Thomas K, Carissa K, Dylan S, Mark H, Brianna C, Wesley M, Austin K, Kieran Mc, Brooke H
  • Date: March 13, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Antibiotic: Amp
  • Part Number and Plasmid: R0010 PSB1A2, B0032 PSB1A2, J04650 PSB1AK3, B0031 PSB1A2, B0030 PSB1A2, J04500 PSB1AK3
  • Results: TBA

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Miniprep and Glycerol Stock

  • Names: Mark H, Chris I, Elaine B, Dax L
  • Date: March 13, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Miniprepped: 3-B0030, 3-B0031, 3-B0032, 2-J04500 and 3-R0010
  • Glycerol Stocked B0030 in G1, B0031 in G2, B0032 in G3, J04500 in G4 and R0010 in G5

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Restriction Digestion

  • Names: Patrick O, Steven T, Amrinder G, Yoyo Y, Wesley M, Orion S
  • Date: March 15, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe use buffer 4 instead of buffer 10
  • Restricted: 6-B0030, 6-B0031, 6-B0032, 4-J04500 and 6-R0010
  • Incubate 1 hour at 37˚ C
  • Stored freezer overnight

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Agarose Gel

  • Name: Patrick O
  • Date: March 16, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Ran 15 uL each of 3-B0030, 3-B0031, 3-B0032, 2-J04500 and 3-R0010 through 1% agarose gel
  • Quantities: 12.5 uL DNA and 2.5 uL of 6x loading dye (15 uL total), 12.5 uL 1kb DNA ladder and 2.5 uL 6x loading dye (15 uL total) note: ladder difficult to see on gel picture, next time less DNA ladder
Lane # Contents Volume(µL)
1 Empty n/a
2 1kb DNA Ladder 15
3 BBa_B0030 15
4 BBa_B0030 15
5 BBa_B0030 15
6 BBa_B0031 15
7 BBa_B0031 15
8 BBa_B0031 15
9 BBa_B0032 15
10 BBa_B0032 15
11 BBa_B0032 15
12 BBa_R0010 15
13 BBa_R0010 15
14 BBa_R0010 15
15 BBa_J04500 15
16 BBa_J04500 15
17+ empty n/a

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Agarose Gel

  • Names: Fiona S, Katie T, Chris I, Elaine B.
  • Date: March 18, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Ran 15 uL each of 3-B0030, 3-B0031, 3-B0032, 2-J04500 and 3-R0010 through 1% agarose gel
  • Quantities: 12.5 uL DNA and 2.5 uL of 6x loading dye (15 uL total), 12.5 uL 1kb DNA ladder and 2.5 uL 6x loading dye (15 uL total)
  • Lane # Contents Volume(µL)
    1 Empty n/a
    2 1 kb DNA Ladder 6
    2 1 kb DNA Ladder 6
    3 BBa_B0030 15
    4 BBa_B0030 15
    5 BBa_B0030 15
    6 BBa_B0031 15
    7 BBa_B0031 15
    8 BBa_B0031 15
    9 BBa_B0032 15
    10 BBa_B0032 15
    11 BBa_B0032 15
    12 BBa_R0010 15
    13 BBa_R0010 15
    14 BBa_R0010 15
    15 BBa_J04500 15
    16 BBa_J04500 15
    17+ Empty n/a

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Transformations of J04650

  • Names: Fiona S, Chris I.
  • Date: March 19, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Transformed 2 uL of J04650 pSB1AK3 from Spring 2012 Kit Plate 4 C12 into competent cells.
  • Quantities: 2 uL J04650 pSB1AK3, 200 uL LB media

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Picking Cells of J04650

  • Names: Yoyo Y, Wesley M
  • Date: March 20, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked cells of J04650 from previous day’s transformation
  • Colonies didn't form properly within either plate, cells were picked with a dab (single colonies)
  • Quantities: 3x5ml of media
  • Antibiotic: Ampicillin
  • Glycerol Stock: R0010 pSB1A2

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Miniprep and Glycerol Stock of R0010

  • Name: Mark H
  • Date: March 21, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Miniprepped 1-R0010 used 50uL of Elution Buffer
  • Glycerol Stocked 1-R0010 in G6

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Transformations J04650 and B0015

  • Name: Yoyo Y
  • Date: March 27, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Transformed 2uL of J04650, and B0015 into 25uL of DH5A cell
  • Quantities: 2uL of J04650 and 2uL of B0015 added into 25uL of DH5A cells

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Transformations

  • Names: Marie C, Fiona S, Chris I
  • Date: March 28, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Transformed 2.5uL of J06702
  • Plasmid: pSB1A2
  • Quantities: 2.5uL of DNA into 100uL of pSB1A2

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Picking Cells of J04650

  • Names: Chris I, Fiona S, Marie C
  • Date: March 28, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked 3 colonies from March 19, J04650
  • Picked 1 colony from March 26, J04650
  • Picked 1 colony from March 27, B0015
  • 5mL cultures with 5uL ampicillin.

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Picking Cells of J06702

  • Names: Chris I, Fiona S, Marie C
  • Date: March 29, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked 2 colonies from March 28, J06702
  • 5mL cultures with 5uL ampicillin

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Picking Cells of -80 Glycerol Stock , J61002, J23113, J23108, J23111, J23100

  • Names: Chris I, Fiona S, Marie C
  • Date: March 29, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked, J61002
  • Picked, J23113
  • Picked, J23108
  • Picked, J23114
  • Picked, J23100
  • 5mL cultures with 5uL ampicillin
  • Shaken overnight at 37 degrees Celsius

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