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Team:Brown/Notebook/June28
From 2010.igem.org
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(→Preparation of Agar Plates) |
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===Preparation of Agar Plates=== | ===Preparation of Agar Plates=== | ||
| - | + | Agar recipe – cook on hot plate | |
| - | + | *10g NaCl | |
| - | + | *10g tryptone | |
| - | + | *5g yeast extract | |
| - | + | *20g agar | |
| - | + | *Deionized water to a final volume of 1L | |
#Autoclave | #Autoclave | ||
#Let cool down, add Ampicillin to 100 µl/ml; mix well. (Add 1 ml amp at 100 mg/ml stock to 1L to get final concentration of 100 µg/ml.) | #Let cool down, add Ampicillin to 100 µl/ml; mix well. (Add 1 ml amp at 100 mg/ml stock to 1L to get final concentration of 100 µg/ml.) | ||
Latest revision as of 19:26, 19 July 2010
Monday, June 28 2010
Competent cells:
Overnight culture (from Sunday night)
Diluted 1:250 in 250 mL LB media
Incubate (shaking) 37°C until ~0.4 OD at 600 nm (3 to 4 hours).
- Started at 10:50 AM
12:55 PM – Transformation of pGEM ligation product into XL1Blues.
Preparation of Agar Plates
Agar recipe – cook on hot plate
- 10g NaCl
- 10g tryptone
- 5g yeast extract
- 20g agar
- Deionized water to a final volume of 1L
- Autoclave
- Let cool down, add Ampicillin to 100 µl/ml; mix well. (Add 1 ml amp at 100 mg/ml stock to 1L to get final concentration of 100 µg/ml.)
- Pour
Received our L. lactis from Ireland. 50 µl pPTPi, MG1614, and pPTPi in MG1614 (can act as a control)
Grown in M17 broth with 0.5% glucose, 6 µg/mL tetracycline at 30°C.
