Notebook
Everyday activities by the team
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July
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Week 1: 9-15
Pre-Interlab: Transformation Practice (p. 25, 07/13/18)
Starting time: 11:45 am
Antibiotic stock solutions (Carlos)
- Ampicillin 100 mg/ml
- Chloramphenicol 35 mg/ml
- Kanamycin 35 mg/ml
LB Medium (without agar) (Jesús)
- 25 g/L, 500 ml of medium
LB Medium (with agar) (Jesús)
- 25 g/L, 500 ml of medium
- 15 g/L of agar, 500 ml of medium
Medium LB and silica spheres are put inside autoclave
- 121°C for 15 minutes
- Materials left inside fume hood, with 15 min. of UV light
- Bain-marie prepared, 42°C
Pre-Interlab: Transformation Practice (p. 25, 07/13/18)
Transformations (Samantha)
- 4 Petris, 23K + antibiotic, 8P + antibiotic, 23K w/o antibiotic, competent w/o antibiotic
- Positive control: 8P, dish 3, kit 2018: BBa_I20270 (GFP)
- 1: 23K, dish 3, kit 2018: BBa_I763007 (RFP)
DNA from plates to Eppendorfs
- 1: 50 μL CC, 1 μL 23K
- 2: 50 μL CC, 1 μL 23K
- 3: 50 μL CC, 1 μL 8P
- 4: 50 μL CC
Plates (Victor)
- 20 ml LB with agar for all plates
- 20 μL of Chloramphenicol when needed
Heat Shock (Samantha)
- 1-4: 1 minute, 42°C, then 5 minutes on ice + 200 μL LB to each tube
Note: iGEM protocol says 450 ml of SOC, but instead we used 200 μL of LB
Incubations
- 37°C and 220 rpm, during 1 hour (6:03-7:03 pm)
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Week 2: 16-22Interlab: Plate 7 Transformations (p. 28-29, 07/17/18-07/18/18) Devices Wells Eppendorf Ctrl(-).BBa_R0040 7-2D, 7-4D, 7-21D, 6-20D, 2-6F (-) Ctrl(+).BBa_I20270 7-2B, 7-4B, 7-21B, 6-20B, 3-8F (+) D1.BBa_J364000 7-2F, 7-4F, 7-21F, 6-20F 1 D2.BBa_J364001 7-2H, 7-4H. 7-21H, 6-20H 2 D3.BBa_J364002 7-2J, 7-4J, 7-21J, 6-20J 3 D4.BBa_J364003 7-2L, 7-4L 4 D5.BBa_J364004 7-2N, 7-4N 5 D6.BBa_J364005 7-2P, 7-4P 6 DNA Resuspension (Samantha) With respect to the wells on plate 7 Competent cells 50 μL to each Eppendorf duplicate Resuspended DNA 2 μL were added to each Eppendorf duplicate Colonies 1.1 1.2 1.3 2.1 2.2 2.3 3.1 3.2 3.3 4.1 4.2 4.3 D3 388 417 381 536 491 523 498 452 575 548 559 300 D4 71 167 213 223 200 197 255 242 165 116 269 281 D5 15 52 61 65 50 218 55 78 50 56 79 97
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August
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Week 1: 1-5This is text for Week 1
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Week 2: 6-12This is text for Week 1
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Week 3: 13-19This is text for Week 1
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Week 4: 20-26This is text for Week 1
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Week 5: 27-31This is text for Week 1
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September
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Week 1: 1-2This is text for Week 1
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Week 2: 3-9This is text for Week 1
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Week 3: 10-16This is text for Week 1
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Week 4: 17-23This is text for Week 1
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Week 5: 24-30This is text for Week 1
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October
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Week 1: 1-7This is text for Week 1
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Week 2: 8-14This is text for Week 1
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Week 3: 15-17This is text for Week 1
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Protocols
Protocol