Difference between revisions of "Team:Calgary/Protocols"

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                             Rehydration of Synthesized DNA
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                             Rehydration of Synthesized DNA
 
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                                     <p>Synthesized DNA from IDT or Genscript</p>                                
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                                     <ul>
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                                    <p>ddH₂O</p>
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                                        <li>Synthesized DNA from IDT or Genscript</li>
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                                        <li>ddH₂O</li>
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                                     <ol>
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                                         <li>Centrifuge tube containing the synthesized DNA for 1 minute at 3000g</li>
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                                         <li>Centrifuge tube containing the synthesized DNA for 1 minute at 3000g.</li>
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                                         <li>Add 20 μL ddH₂O</li>
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                                         <li>Add 20μL ddH₂O.</li>
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                                         <li>Vortex for 1 minute</li>
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                                         <li>Vortex for 1 minute.</li>
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                                         <li>Incubate at 50°C for 15 minutes</li>
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                                         <li>Incubate at 50°C for 15 minutes.</li>
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                                       <li>Briefly centrifuge. Store at -20°C</li>
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                                       <li>Briefly centrifuge. Store at -20°C.</li>
 
                                     </ol>
 
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Revision as of 23:16, 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:
    1. 10% (w/v) tryptone
    2. 5% (w/v) NaCl
    3. 10% (w/v) yeast extract
    4. 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.