Difference between revisions of "Team:Marburg/Labbook/Minicells"
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| − | Cultures containing a plasmid which encodes enzymes for the biosynthesis of β-carotene were treated according to the Macherey-Nagel Plasmid DNA Purification Kit protocol. | + | Cultures containing a plasmid which encodes enzymes for the biosynthesis of β-carotene (Car) were treated according to the Macherey-Nagel Plasmid DNA Purification Kit protocol. |
</p> | </p> | ||
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</p> | </p> | ||
| + | <h1>15/08/29</h1> | ||
| + | <h2>Preparation of ONC</h2> | ||
| + | <p> | ||
| + | Four times 5 mL LB was inoculated with TB43 Car and incubated at 37 °C and 200 rpm. | ||
| + | </p> | ||
| + | <h1>15/08/30</h1> | ||
| + | <h2>Plasmid Preparation</h2> | ||
| + | |||
| + | <p> | ||
| + | ONC were treated according to the Macherey-Nagel Plasmid DNA Purification Kit protocol. | ||
| + | </p> | ||
| + | |||
| + | <h1>15/09/03</h1> | ||
| + | |||
| + | <h2>Preparation of ONC</h2> | ||
| + | |||
| + | <p> | ||
| + | 5 mL LB were inoculated with TB43, TB43 GFP, TB43 HU-mCherry, TB43 HU-mCherry GFP and TB43 Car and incubated at 37 °C and 200 rpm. | ||
| + | </p> | ||
| + | |||
| + | <h1>15/09/04</h1> | ||
| + | |||
| + | <h2>Preparation of ONC</h2> | ||
| + | |||
| + | <p> | ||
| + | 5 mL M9 minimal medium were inoculated with TB43, TB43 GFP, TB43 HU-mCherry, TB43 HU-mCherry GFP and TB43 Car and incubated at 37 °C and 200 rpm. | ||
| + | </p> | ||
| + | |||
| + | <h1>15/09/05</h1> | ||
| + | |||
| + | <h2>Growth Curve</h2> | ||
| + | |||
| + | <p> | ||
| + | TB43, TB43 HU-mCherry, TB43 GFP and TB43 HU-mCherry GFP were cultivated in LB and M9 minimal medium. No minicells could be detected by flow cytometry. | ||
| + | </p> | ||
| + | |||
| + | <h2>Preparation of ONC</h2> | ||
| + | |||
| + | <p> | ||
| + | 5 mL | ||
Revision as of 19:32, 11 September 2015
15/07/07
Preparation of ONCs
Two times 5 mL LB were inoculated with TB43 and TB43 HU-mCherry and incubated at 37 °C and 200 rpm.
15/07/08
Preparation of Glycerol Stocks
Glycerol stocks of TB43 and TB43 HU-mCherry were prepared according to the glycerol stock generation protocol.
Preparation of electrocompetent Cells
Electrocompetent cells of TB43 and TB43 HU-mCherry were prepared according to the electrocompetent E.coli cells protocol.
Preparation of over-night culture (ONC)
Two times 5 mL LB were inoculated with TB43 and TB43 HU-mCherry and incubated at 37 °C and 200 rpm.
15/07/09
Preparation of electrocompetent Cells
Electrocompetent cells of TB43 and TB43 HU-mCherry were prepared according to the electrocompetent E.coli cells protocol.
15/07/10
Preparation of day culture
20 mL LB were inoculated with TB43 and TB43 HU-mCherry for flow cytometry and grown to an optical density (OD) of 0.5. No minicells could be detected.
15/07/11
Transformation
A GFP plasmid with Chloramphenicol resistance gene was transformed into TB43 and TB43 HU-mCherry via electroporation according to transformation into electrocompetent cells protocol.
15/07/13
Preparation of ONC
50 mL LB were inoculated with TB43 GFP and TB43 HU-mCherry GFP and incubated at 37 °C and 200 rpm.
15/07/15
Preparation of Minicells
TB43 GFP and TB43 HU-mCherry GFP were treated according to preparation of minicells protocol. Samples were taken and minicells could be detected via flow cytometry but only with very low yields.
15/07/27
Preparation of ONC
5 mL LB were inoculated with TB43, TB43 HU-mCherry, TB43 GFP and TB43 HU-mCherry GFP and incubated at 37 °C and 200 rpm.
15/07/28
Preparation of chemocompetent Cells
Chemocompetent cells of TB43, TB43 GFP, TB43 HU-mCherry and TB43 HU-mCherry GFP were prepared according to competent E.coli cells (RbCl) protocol.
15/08/21
Preparation of ONC
5 mL LB were inoculated with TB43, TB43 HU-mCherry, TB43 GFP and TB43 HU-mCherry GFP and incubated at 37 °C and 200 rpm.
Preparation of M9 Minimal Medium
Two media were prepared. One according to M9 minimal medium protocol and the other without glucose.
15/08/24
Plasmid Preparation
Cultures containing a plasmid which encodes enzymes for the biosynthesis of β-carotene (Car) were treated according to the Macherey-Nagel Plasmid DNA Purification Kit protocol.
15/08/25
Transformation
Carotinoid plasmids were transformed into chemocompetent TB43 and TB43 HU-mCherry according to Transformation into chemocompetent (RbCl) Cells protocol.
Plasmid Preparation
Cultures containing a plasmid which encodes enzymes for the biosynthesis of violacein were treated according to the Macherey-Nagel Plasmid DNA Purification Kit protocol.
15/08/29
Preparation of ONC
Four times 5 mL LB was inoculated with TB43 Car and incubated at 37 °C and 200 rpm.
15/08/30
Plasmid Preparation
ONC were treated according to the Macherey-Nagel Plasmid DNA Purification Kit protocol.
15/09/03
Preparation of ONC
5 mL LB were inoculated with TB43, TB43 GFP, TB43 HU-mCherry, TB43 HU-mCherry GFP and TB43 Car and incubated at 37 °C and 200 rpm.
15/09/04
Preparation of ONC
5 mL M9 minimal medium were inoculated with TB43, TB43 GFP, TB43 HU-mCherry, TB43 HU-mCherry GFP and TB43 Car and incubated at 37 °C and 200 rpm.
15/09/05
Growth Curve
TB43, TB43 HU-mCherry, TB43 GFP and TB43 HU-mCherry GFP were cultivated in LB and M9 minimal medium. No minicells could be detected by flow cytometry.
Preparation of ONC
5 mL
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