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Team:Brown/Notebook/June28
From 2010.igem.org
(Difference between revisions)
(New page: {{:Team:Brown/templates/header}} ==Monday, June 28 2010== ===Competent cells:=== Overnight culture (from Sunday night) Diluted 1:250 in 250 mL LB media Incubate (shaking) 37°C until ...) |
(→Preparation of Agar Plates) |
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#Agar recipe – cook on hot plate. | #Agar recipe – cook on hot plate. | ||
| - | *10g NaCl | + | **10g NaCl |
| - | *10g tryptone | + | **10g tryptone |
| - | *5g yeast extract | + | **5g yeast extract |
| - | *20g agar | + | **20g agar |
| - | *Deionized water to a final volume of 1L | + | **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.) | ||
Revision as of 19:21, 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.
