Interlab protocol

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

(see https://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