Team:Goettingen/Experiments



Media

LB Medium

Phosphatase Activity plates, Sperber media

Esterase Activity plates, with 1% Tributyrin

Cellulase activity plates

Cloning Methods

PCR product purification using QIAquick® PCR Purification Kit (QIAGEN)

PCR Gel extraction, peqGOLD Gel Extraction Kit

Blunt End Ligation in pJET1.2 vector –Clone JET PCR Cloning Kit–Thermo Scientific

TOPO® Cloning protocol usingChampion™ pET Directional TOPO® Expression Kits. Thermo Fisher Scientific

Plasmid transformation into chemically competent E. coli

Electroporation of BL21 cells with pJET_RFP

Plasmid Extraction - using QIAprep Spin Miniprep Kit (QIAGEN)

Plasmid Extraction - using peqGOLD Plasmid Miniprep Kit I (PEQLAB Technologies)

Competent Cells

Preparation of competent E.coli cells

Competent Cell Test Kit, RFP construct (iGEM)

Activity Screens

Esterase activity test

Cellulase activity screening

Restriction controls

Aan I (Psi I ) - thermo fisher scientific - restriction control protocol

Double digestion restriction control

Restriction control using fast and slow digestion enzymes

Scafoldin Restriction control

Esterase Restriction Control

Phosphatase Restriction Control

PCR preparation methods

Colony PCR

Phusion PCR

Sequencing

Protocol for Sanger sequencing

Sanger sequencing reaction setup for overnight