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 [http://openwetware.org/wiki/Bacterial_cell_culture 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 [http://openwetware.org/wiki/Knight:Colony_PCR Knight - Colony PCR]

NEB Taq

Finnzymes Phusion

DNA Extraction

Quiagen Miniprep Isopropanol Extraction of small DNA fragments

Construction of BioBricks

Gingko Bioworks - [http://ginkgobioworks.com/support/ Three Antibiotic Assembly (NEB Biobrick Assembly Kit)]

OpenWetWare [http://openwetware.org/wiki/In-fusion_biobrick_assembly In Fusion Biobrick Assembly (Primer design and assembly strategy)]

Clontech [http://www.clontech.com/products/detail.asp?catalog_id=639621 In Fusion Cloning (In Fusion reaction itself)]

OpenWetWare [http://openwetware.org/wiki/Endy:Annealing_complementary_primers Endy - Annealing Complementary Primers]