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Team:ETH Zurich/labblog/20140703
From 2014.igem.org
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| - | + | == Sequencing, Transformation, antibiotic stock preparation == | |
| - | + | = Thursday, July 3rd = | |
| + | |||
| + | <html><section class="frag"></html> | ||
| + | ===Sequencing results=== | ||
| + | Biobricks of the plasmids piG0001, piG0015 and piG002 show the correct sequences | ||
<html></section></html> | <html></section></html> | ||
| + | |||
| + | <html><section class="frag"></html> | ||
| + | ===E coli Transformation=== | ||
| + | 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 | ||
| + | Heat shock: 90 s at 42 °C<br/> | ||
| + | Addition of SOC to samples, let cells recover for ca. 1h at 37 °C, 220 rpm<br/> | ||
| + | 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))<br/> | ||
| + | Let bacteria grow overnight at 37 °C | ||
| + | |||
| + | <html></section></html> | ||
| + | |||
| + | <html><section class="frag"></html> | ||
| + | |||
| + | ===Antibiotic stock preparation=== | ||
| + | Preparation of antibiotics stock:<br/> | ||
| + | 2 g Ampicillin were dissolved in 10 mL H2O and sterile filtered <br/> | ||
| + | 0.5 g Kanamycin were dissolved in 10 mL H2O and sterile filtered <br/> | ||
| + | 0.34 g Chloramphenicol were dissolved in 10 mL Ethanol<br/> | ||
| + | <html></section></html> | ||
| + | |||
| + | [[#top|Top]] | ||
| + | <html></article></html> | ||
Latest revision as of 09:51, 10 July 2014
Contents |
Sequencing, Transformation, antibiotic stock preparation
Thursday, July 3rd
Sequencing results
Biobricks of the plasmids piG0001, piG0015 and piG002 show the correct sequences
E coli 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
Antibiotic stock preparation
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
