Difference between revisions of "Team:Tec-Monterrey/Notebook"

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

 

 

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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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Interlab: 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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