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Team:Lethbridge Canada/notebook march
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<li><a href="https://2013hs.igem.org/Team:Lethbridge_Canada/outreach">Overview</a></li> | <li><a href="https://2013hs.igem.org/Team:Lethbridge_Canada/outreach">Overview</a></li> | ||
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Revision as of 22:35, 14 June 2013
Notebook Links
- March
- March 12, 2013: Transformation
- March 13, 2013: Picking Cell Colonies (Cultures)
- March 13, 2013: Miniprep and Glycerol Stock
- March 15, 2013: Restriction Digestion
- March 16, 2013: Agarose Gel
- March 18, 2013: Agarose Gel
- March 19, 2013: Transformation
- March 20, 2013: Picking Cell Colonies (Cultures)
- March 21, 2013: Miniprep and Glycerol Stock
- March 27, 2013: Transformation
- March 28, 2013: Transformation
- March 28, 2013: Picking Cell Colonies (Cultures)
- March 29, 2013: Picking Cell Colonies (Cultures)
- March 29, 2013: Picking Cells of -80 Glycerol Stock
- April
- April 2, 2013: Restriction - Ligation
- April 2, 2013: Ligation
- April 3, 2013: Transformation
- April 4, 2013: Re-Plating Transformation
- April 5, 2013: Miniprep of ON Cultures
- April 5, 2013: UV Spectroscopy DNA Concentrations
- April 6, 2013: Miniprep From Ligations
- April 7, 2013: PCR
- April 8, 2013: Restriction Digestion of PCR Parts
- April 8, 2013: 3% Agarose Gel of Digested PCR Parts
- April 8, 2013: Gel Extraction
- April 9, 2013: Repeat PCR
- April 9, 2013: Picking Cell Colonies (Cultures)
- April 10, 2013: Miniprep
- April 10, 2013: Restriction
- April 10, 2013: Agarose Gel of PCR 1%
- April 11, 2013: Miniprep
- April 12, 2013: Restriction Digestion
- April 12, 2013: PCR and Cell Reculturization
- April 12, 2013: Miniprep
- April 14, 2013: Miniprep
- April 14, 2013: 1% Agarose Gel
- April 15, 2013: Restriction/Digestion
- April 15, 2013: Restriction/Digestion
- April 15, 2013: 1% Agarose Gel of Promoters
- April 17, 2013: Transformation
- April 30, 2013: 1% Agarose Gel of Promoter Constructs
- May
- May 1, 2013: Ligation
- May 4, 2013: Restriction Digest
- May 4, 2013: 1% Agarose Gel of Promoter Constructs
- May 4, 2013: Gel Extractions
- May 5, 2013: Transformation
- May 7, 2013: Restriction
- May 7, 2013: Gel Extraction
- May 7, 2013: Gel Extracted on Conformation Gel
- May 8, 2013: Ligations
- May 9, 2013: PCR of Ligation Mixture
- May 13, 2013: Restriction of PCR Ligations Gel Conformation
- May 14, 2013: Transformation of Ligations
- May 15, 2013: Ligation
- May 15, 2013: Picking Cells
- May 16, 2013: Picking Cells
- May 16, 2013: 10 Fold Miniprep
- May 17, 2013: Agarose Gel
- May 17, 2013: Miniprep
- May 17, 2013: Glycerol Stock
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
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
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
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
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 |
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
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
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
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
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
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
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.
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
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
