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

[http://www.neb.com/nebecomm/products/protocolProductC2984.asp NEB Turbo Competent E. coli]

[http://www.clontech.com/products/detail.asp?product_id=215795&tabno=3 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

[http://www.qiagen.com/products/plasmid/qiaprepminiprepsystem/qiaprepspinminiprepkit.aspx#Tabs=t2 Quiagen Miniprep]

Isopropanol Extraction of small DNA fragments

Construction of BioBricks

Gingko Bioworks - [http://ginkgobioworks.com/support/ 3A Biobrick 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]