Difference between revisions of "Team:Calgary/Protocols"

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                                     <p>Luria-Bertani broth with agar:</p>
                                        <li>Luria-Bertani broth with agar:</li>
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                                       <ol>
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                                               <li>10% (w/v) tryptone</li>
 
                                               <li>10% (w/v) tryptone</li>
 
                                               <li>5% (w/v) NaCl</li>
 
                                               <li>5% (w/v) NaCl</li>
 
                                               <li>10% (w/v) yeast extract</li>
 
                                               <li>10% (w/v) yeast extract</li>
 
                                               <li>15% (w/v) agar</li>
 
                                               <li>15% (w/v) agar</li>
                                   </ol>
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                                   </ul>
                                        <li>Appropriate antibiotic:</li>
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                                    <p>Appropriate antibiotic:</p>                                  
                                    </ul>
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Revision as of 23:22, 15 October 2018

Team:Calgary/Notebook - 2018.igem.org

PROTOCOLS




Below are the protocols used by the team.

Materials

iGEM 2018 distribution kit

  • List Item 1
  • List Item 2

ddH₂O

Protocol
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA as per transformation protocol. Store the remaining amount at -20°C.

Materials

Synthesized DNA from IDT or Genscript

ddH₂O

Protocol
  1. Centrifuge tube containing the synthesized DNA for 1 minute at 3000g.
  2. Add 20μL ddH₂O.
  3. Vortex for 1 minute.
  4. Incubate at 50°C for 15 minutes.
  5. Briefly centrifuge. Store at -20°C.

Materials

Luria-Bertani broth with agar:

  • 10% (w/v) tryptone
  • 5% (w/v) NaCl
  • 10% (w/v) yeast extract
  • 15% (w/v) agar

Appropriate antibiotic:

Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.

Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.

Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.

Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.