IGEM:IMPERIAL/2009/Assays Protocols/Shopping

Module 1

Cellulase

Link to Assay

Reagents:
Sodium Acetate
Sigmacell Solution
Cellulase Enzyme Solution
Glucose Determination Vial

PAH

Link to Assay

Reagents:
Tris CH1 Buffer
L-Phenylalanine Solution
Catalase Enzyme Solution
DL-Dithiothreitol Solution
6-Methyltetrahydropterine Solution

Module 2

Colanic Acid

Link to Assay Page
Protocol for Quantitative Assay
Qualitative Assay using electron microscopy could be performed as well, to ensure sufficient encapsulation.

Reagents:
No special reagents needed. Centrifuge performed on bacterial media.

Equipment:
PCV Centrifuge Tubes

OtsA

Link to Assay

Trehalose 6-Phosphate Synthases Catalyses the following reaction:
GDP-glucose + glucose 6-phosphate --> GDP + alpha,alpha-trehalose 6-phosphate

We will perform the quantitative assay shown above to assess the activity of T6PS

Reagents Required: UDPG
G-6-P
Tris-HCl Buffer
MnCl2
heparin salt
HCl
NaOH
anthrone
Shodex Sugar

OtsB (Trehalose) Assay

Link to Assayys Purchasing Page

Trehalose 6-Phosphate Phosphatase catalyses the following reaction:

alpha,alpha-trehalose 6-phosphate + H2O --> alpha,alpha-trehalose + phosphate

We will therefore perform a trehalose assay on the products of OtsB to see if functional T6PP is produced. This will give us a quantifiable output to the POPs input to the gene.