Team:St Andrews/project/laboratory/protocols

Contents 1 Protocols 1.1 Cell Culture 1.2 Transformation 1.3 Restriction Digest 1.4 Ligation 1.5 PCR 1.6 DNA Extraction 1.7 Construction of BioBricks

Protocols

We used a number of protocols from OpenWetWare in addition to the manufacturer's recommended protocols for our materials.

Cell Culture

OpenWetWare Bacterial Cell Culture

Transformation

NEB Turbo Competent E. coli

Clontech Stellar Competent E. coli.

Restriction Digest

Promega Enzymes

NEB Biobrick Assembly Kit (Gingko Bioworks)

Ligation

NEB Biobrick Assembly Kit (Gingko Bioworks)

PCR

OpenWetWare Knight - Colony PCR

NEB Taq

Finnzymes Phusion

DNA Extraction

Quiagen Miniprep

Isopropanol Extraction of small DNA fragments

Construction of BioBricks

Gingko Bioworks - 3A Biobrick Assembly (NEB Biobrick Assembly Kit)

OpenWetWare In Fusion Biobrick Assembly (Primer design and assembly strategy)

Clontech In Fusion Cloning (In Fusion reaction itself)

OpenWetWare Endy - Annealing Complementary Primers