Team:Evry/Interlab/Protocol

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Let's PLAy project - Bioproduction of PLA

Interlab protocol


The majority of the protocols for the Interlab study were provided by the iGEM HQ.

(see http://2016.igem.org/Tracks/Measurement/Interlab_study/Plate_Reader_Overview).


Preparation of phosphate buffered saline 1X solution
  • 8g NaCl for final concentration [137 mM]
  • 0.2 KCl [2.7 mM]
  • 1.44g Na2HPO4 [1 mM]
  • 0.24g KH2PO4 [1.8 mM]

Add 800µL dH20 and mix. Then, adjust pH= 7.4 (7.2 if necessary) with HCl.

When the pH is good, qs 1 L. 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

9.9 mL

Sorbitol 5X (2.5M)

2 mL

Betaine 100X (500 mM)

100 u