08/08/13

From 2013.igem.org

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