Team:Stuttgart/Experiments

Experiments and methods

Media and agar

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LB Agar for E.coli

LB Medium for E.coli

Low salt LB

SOC medium

P. pastoris culture medium

1M Potassium phosphate buffer pH 6

1M Sorbitol

10X Dextrose

10X Glycerol

10X YNB

Media 500X Biotin

Media 500X Cu₂SO₄

Minimal dextrose medium (MD)

Minimal methanol medium (MM)

Stock BMGY and BMMY

Stock YPD

Stock YPDS agar

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Cloning

Bacterial laccase gene

Competent cells and transformation

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Generation of chemical competent cells

Transformation

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Cloning

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PCR for amplification

DNA Assembly chemical

DNA Assembly electrocompetent

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DNA Assembly Reaction Protocol

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Ligation of pET22b and modified laccase of S. cyaneus

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Restriction digest of pET22b and modified laccase of S. cyaneus

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Yeast laccase gene

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PCR for amplification

Mutant selection

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Laccase Production

Prodcution of bacterial laccase

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Transformation for protein purification

Protein expression

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Production of yeast laccase

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Generation of competent cells (P.pastoris)

Transforamtion via electroporation

Protein expression in P.pastoris

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Mesocellular silicafoam (MCF)

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MCF Synthesis

MCF Immobilization

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Assays

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ABTS assay

Bradford assay

Diclofenac and carbamzepine assay

HPLC method

Longterm stability of T.versicolor

pH stability assay

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© Christopher Adelmann, Fabian Klötzer, Jan Schlegel for iGEM Team Stuttgart 2020