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Protocol

http://ginkgobioworks.com/support/

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OpenWetWare Protocols: http://openwetware.org/wiki/Protocols

Contents 1 DNA (well to stock) 2 Bacterial Work 2.1 Transformation 2.2 O/N Culture 2.3 Preparation of Competent Cells 2.4 LB Media 2.5 Preparing Plates 3 DNA 3.1 Restriction Enzyme Digests 3.2 Ligations 3.3 Mini Prep

DNA (well to stock)

1. With a pipette tip, punch a hole through the foil cover into the corresponding well to the Biobrick™-standard part that you want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to cross contamination between the wells.
2. Add 10uL of diH2O (deionized water)
3. Pipette 1 or 2uL of the resuspended DNA transform into your desired competent cells, plate bacteria with the appropriate antibiotic* and grow overnight.
4. Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 18 hours.
5. Use the resulting culture to miniprep the DNA AND make your own glycerol stock (for further instruction on making a glycerol see this page). We recommend using the miniprepped DNA to run QC tests, such as restriction digests and sequencing.

Bacterial Work

Transformation

O/N Culture

Preparation of Competent Cells

LB Media

Preparing Plates

DNA

Restriction Enzyme Digests

Ligations

Mini Prep