Difference between revisions of "Team:DTU-Denmark/Experiments"

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Minimal medium: Schizophyllum commune SMM agar (1L) needed for S.commune plates
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<h4 class="media heading">Materials</h4>
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  <li><p>Glucose - 20g</p></li>
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<li><p>Monopotassium phosphate KH<sub>2<\sub>PO<sub>4</sub> - 0.46g</p></li>
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<li><p>Potassium phosphate dibasic K<sub>2<\sub>HPO<sub>4<\sub>  3H<sub>2<\sub>O - 1.28g</p></li>
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<li><p>Magnesium sulfate MgSO<sub>4<\sub>  7H<sub>2<\sub>O - 0.5g</p></li>
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<li><p>Trace elements solution - 1ml </p></li>
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<li><p>FeCl<sub>3<\sub> solution - 1ml</p></li>
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<li><p>L- Asparagin - 1.5g</p></li>
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<li><p>Agar - 20g</p></li>
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<li><p>Thiamine (10mg/100ml) - 1.2ml </p></li>
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<h4 class="media heading">Procedure</h4>
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<p>Preparation of the media
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  <li><p>Weigh and add the required solid components (except agar)</p></li>
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<li><p>Add the required liquid components in a bottle</p></li>
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<li><p>Dissolve in demineralized water until the total volume is 900ml </p></li>
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  <li><p>Stir the liquid using a magnet (no pH adjustment is required)</p></li>
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<li><p>Add the required quantity of agar </p></li>
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<li><p>Add demineralized water until the desired total volume (1L) </p></li>
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  <li><p>Autoclave </p></li>
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<li><p>After sterilization add 1.2 ml of filter sterilized thiamine</p></li>
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Revision as of 16:52, 4 October 2018

Experiments

Synthlab protocols

DNA from IDT will typically be delivered in a white flaky substance, which need to be resuspended, before the DNA is ready for use

Materials

  • Table centrifuge

  • EB/TE buffer

  • Genes/Primers from IDT or other DNA provider

Procedure

  1. Quickly spin the DNA down in the table centrifuge

  2. Calculate the amount of EB buffer need to dilute the gblocks to a desired concentration. Important: gene fragments and primeres are not diluted to the same concentration: The concentration of gene fragments is usually 25 ng/μL and for primers it is 100 μM.

  3. Genes/Primers from IDT or other DNA provider

    • DNA from IDT usually comes in dried flakes of 500 or 1000 ng of DNA. To achieve the desired concentration (usually 25 ng/μL) the needed amount of EB buffer is 40 μL (for 1000 ng samples) or 20 μL (for 500 ng samples)

    • In order to calculate the the molar amount of primer, use the NEB calculator

  4. Add the calculated amount of EB buffer

  5. Store the resuspended DNA at -20°C

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.

Mycolab protocols

Basic protocol on plating of the fungi.

Materials

  • Agar plates with media of interest

  • Sterile toothpicks

Procedure

Plating using mycelia

  1. Open the plate containing the mycelia from the species of interest

  2. With a sterile toothpick, scratch the surface of the mycelia.

  3. Spike the new agar plate with the toothpick.

  4. Repeat steps 1-3 times. Three inoculation points should be made in the agar making forming a triangle.

  5. Put the plate into a plastic growth bag.

  6. Place growing bag into the incubator.

Plating using spores

  1. Touch the sterile tooth pick in the spore suspension.

  2. Spike the new agar plate with the toothpick.

  3. Repeat steps 1-3 times. Three inoculation points should be made in the agar making forming a triangle.

  4. Put the plate into a plastic growth bag.

  5. Place growing bag into the incubator.

Minimal media used for protoplastation of A. oryzae. This is for 1 L of media.

Materials

  • 50 mL D-glucose

  • 50 mL Nitrate salts

  • 1 mL Trace elements

  • 1 mL Thiamine

  • 20 g Agar (SO.BI.GEL)

Procedure

Mix

  1. Mix in a blue cap flask.

  2. Add water until the volume is 1 L.

  3. Autoclave.

Minimal medium: Schizophyllum commune SMM agar (1L) needed for S.commune plates

Materials

  • Glucose - 20g

  • Monopotassium phosphate KH2<\sub>PO4 - 0.46g

  • Potassium phosphate dibasic K2<\sub>HPO4<\sub> 3H2<\sub>O - 1.28g

  • Magnesium sulfate MgSO4<\sub> 7H2<\sub>O - 0.5g

  • Trace elements solution - 1ml

  • FeCl3<\sub> solution - 1ml

  • L- Asparagin - 1.5g

  • Agar - 20g

  • Thiamine (10mg/100ml) - 1.2ml

Procedure

Preparation of the media

  1. Weigh and add the required solid components (except agar)

  2. Add the required liquid components in a bottle

  3. Dissolve in demineralized water until the total volume is 900ml

  4. Stir the liquid using a magnet (no pH adjustment is required)

  5. Add the required quantity of agar

  6. Add demineralized water until the desired total volume (1L)

  7. Autoclave

  8. After sterilization add 1.2 ml of filter sterilized thiamine