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Team:ETH Zurich/labblog/20140703
From 2014.igem.org
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Transformation of E. coli with pSEVA181, pSEVA271, pSEVA291, pSEVA561.<br/> | Transformation of E. coli with pSEVA181, pSEVA271, pSEVA291, pSEVA561.<br/> | ||
Addition of 1 μL DNA to 75 μL competent cells (thawed on ice), put samples on ice for approximately 20 min | Addition of 1 μL DNA to 75 μL competent cells (thawed on ice), put samples on ice for approximately 20 min | ||
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Preparation of antibiotics stock:<br/> | Preparation of antibiotics stock:<br/> | ||
Revision as of 09:39, 10 July 2014
Contents |
E Coli transformation and antibiotic stock preparation
Thursday, July 3rd
Transformation
Transformation of E. coli with pSEVA181, pSEVA271, pSEVA291, pSEVA561.
Addition of 1 μL DNA to 75 μL competent cells (thawed on ice), put samples on ice for approximately 20 min
Heat shock: 90 s at 42 °C
Addition of SOC to samples, let cells recover for ca. 1h at 37 °C, 220 rpm
Plate 100 μL of bacteria suspension on LB-agar-plates containing the appropriate antibiotics (pSEVA181: Ampicillin (200 μg/L), pSEVA271 and pSEVA291: Kanamycin (50 μg/L), pSEVA561: Tetracycline (20 μg/L))
Let bacteria grow overnight at 37 °C
Title of one section
Preparation of antibiotics stock:
2 g Ampicillin were dissolved in 10 mL H2O and sterile filtered
0.5 g Kanamycin were dissolved in 10 mL H2O and sterile filtered
0.34 g Chloramphenicol were dissolved in 10 mL Ethanol
