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08/08/13
From 2013.igem.org
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| - | ==Making | + | |
| + | ==Making samples for DNA sequencing== | ||
| + | |||
| + | ==Making chloroamphenicol/IPTG plates== | ||
*400ml of agar | *400ml of agar | ||
**Boil for 3min x2 | **Boil for 3min x2 | ||
**Boil for 2min x2 | **Boil for 2min x2 | ||
*Cool to 40C | *Cool to 40C | ||
| - | *Add 800ul of chloroamphenicol at | + | *Add 800ul of chloroamphenicol at 25ug/ml |
| - | *Add 400ul IPTG at 0. | + | *Add 400ul IPTG at 0.15mM |
| + | *Makes 20 petri dishes | ||
| + | |||
| + | ==Streaked out samples of the limonene/pSB1C3 ligated plasmid== | ||
| + | *Streaked samples 5.1, 10.1, 10.2 and 10.3 onto the chloroamphenicol/IPTG plates | ||
| + | *IPTG was used to induce the lac operon, and therefore induce limonene synthesis | ||
| + | *Plates were labelled using special symbols by only one team member to ensure double-blinded test | ||
| + | |||
| + | ==Made polystyrene strips== | ||
| + | --> include video here | ||
| + | |||
| + | ==Overnight culture preparation== | ||
| + | *To make glycerol broths, so sample can be frozen and preserved | ||
| + | *Use four universal tubes, add: | ||
| + | **4ml Luria Broth | ||
| + | **8ul chloroamphenicol | ||
| + | **Add bacteria from the original plates (sample 5.1, 10.1, 10.2 and 10.3 into a single tube each respectively) | ||
| + | |||
| + | *Each sample was also re-streaked on a chloroamphenicol plate to maintain current stock. | ||
Latest revision as of 10:20, 2 September 2013
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Contents |
Making samples for DNA sequencing
Making chloroamphenicol/IPTG plates
- 400ml of agar
- Boil for 3min x2
- Boil for 2min x2
- Cool to 40C
- Add 800ul of chloroamphenicol at 25ug/ml
- Add 400ul IPTG at 0.15mM
- Makes 20 petri dishes
Streaked out samples of the limonene/pSB1C3 ligated plasmid
- Streaked samples 5.1, 10.1, 10.2 and 10.3 onto the chloroamphenicol/IPTG plates
- IPTG was used to induce the lac operon, and therefore induce limonene synthesis
- Plates were labelled using special symbols by only one team member to ensure double-blinded test
Made polystyrene strips
--> include video here
Overnight culture preparation
- To make glycerol broths, so sample can be frozen and preserved
- Use four universal tubes, add:
- 4ml Luria Broth
- 8ul chloroamphenicol
- Add bacteria from the original plates (sample 5.1, 10.1, 10.2 and 10.3 into a single tube each respectively)
