Difference between revisions of "Team:Evry/Interlab/Protocol"
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<h1 class="blue">Interlab protocol</h1> | <h1 class="blue">Interlab protocol</h1> | ||
| − | <p>The main of the protocol for this interlab study was given by the iGEM HQ (https://2016.igem.org/Tracks/Measurement/Interlab_study/Plate_Reader_Overview).</p> | + | <p>The main of the protocol for this interlab study was given by the iGEM HQ (<a href="https://2016.igem.org/Tracks/Measurement/Interlab_study/Plate_Reader_Overview">https://2016.igem.org/Tracks/Measurement/Interlab_study/Plate_Reader_Overview</a>).</p> |
<p> </p> | <p> </p> | ||
<ul> | <ul> | ||
| − | <li><strong>Preparation of phosphate buffered saline 1X solution </strong></li> | + | <li><strong><b>Preparation of phosphate buffered saline 1X solution </b></strong></li> |
</ul> | </ul> | ||
<p style="padding-left: 60px;">*8g NaCl for final concentration [137mM]</p> | <p style="padding-left: 60px;">*8g NaCl for final concentration [137mM]</p> | ||
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<p> </p> | <p> </p> | ||
<ul> | <ul> | ||
| − | <li><strong>Preparation of potassium phosphate solution</strong></li> | + | <li><strong><b>Preparation of potassium phosphate solution</b></strong></li> |
</ul> | </ul> | ||
<p>It is a solution of 0.17 M KH<sub>2</sub>PO<sub>4</sub>, 0.72 M K<sub>2</sub>HPO<sub>4</sub>. Dissolving 2.31 g of KH<sub>2</sub>PO<sub>4</sub> and 12.54 g of K<sub>2</sub>HPO<sub>4</sub> in 90 mL of H<sub>2</sub>O.</p> | <p>It is a solution of 0.17 M KH<sub>2</sub>PO<sub>4</sub>, 0.72 M K<sub>2</sub>HPO<sub>4</sub>. Dissolving 2.31 g of KH<sub>2</sub>PO<sub>4</sub> and 12.54 g of K<sub>2</sub>HPO<sub>4</sub> in 90 mL of H<sub>2</sub>O.</p> | ||
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<p> </p> | <p> </p> | ||
<ul> | <ul> | ||
| − | <li><strong>Preparation of TB medium with betaine + sorbitol </strong></li> | + | <li><strong><b>Preparation of TB medium with betaine + sorbitol </b></strong></li> |
</ul> | </ul> | ||
<p> </p> | <p> </p> | ||
Revision as of 02:11, 20 October 2016
Interlab protocol
The main of the protocol for this interlab study was given by the iGEM HQ (https://2016.igem.org/Tracks/Measurement/Interlab_study/Plate_Reader_Overview).
- Preparation of phosphate buffered saline 1X solution
*8g NaCl for final concentration [137mM]
*0.2 KCl [2.7mM]
*1.44g Na2HPO4 [1mM]
*0.24g KH2PO4 [1.8mM]
Add 800µL dH20 and mix. Then, adjust pH=7.4 (7.2 if necessary) with HCl.
When the pH is good, qs 1L. Sterilize by autoclaving.
- Preparation of 1x TB “Step1”
*Deionized H2O, to 900 mL
*Tryptone, 12 g
*Yeast extract, 24 g
*Glycerol, 4 mL
Shake until the solutes have dissolved and then sterilize by autoclaving for 20 min at 15 psi (1.05 kg/cm2) on liquid cycle.
- Preparation of 1x TB “Ready to use”
Add 100 mL of a sterile potassium phosphate solution in one liter of 1x TB “Step1”.
- Preparation of potassium phosphate solution
It is a solution of 0.17 M KH2PO4, 0.72 M K2HPO4. Dissolving 2.31 g of KH2PO4 and 12.54 g of K2HPO4 in 90 mL of H2O.
After the salts have dissolved, adjust the volume of the solution to 100 mL with H2O and sterilize by autoclaving for 20 min at 15 psi [1.05 kg/cm2] on liquid cycle.
- Preparation of TB medium with betaine + sorbitol
|
|
for 12 mL |
|
TB + RhoPhosphate |
9.9mL |
|
Sorbitol 5X (2.5M) |
2 mL |
|
Betaine 100X (500mM) |
100 u |
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