Experiments

Procedures & Protocols

Purpose

This year, the UT Austin iGEM team worked with Acinetobacter baylyi ADP1 as our primary chassis organism. Handling ADP1 in the lab requires a unique set of protocols that are designed to take advantage of its natural competence. These unique protocols can be used alongside standard molecular biology procedures including PCR, Golden Gate Assembly, and plating techniques, to efficiently engineer ADP1. The experiments below describe both ADP1-specific protocols and standard molecular biology protocols needed to work with ADP1. We hope that providing the iGEM community with these procedures will promote the usage of ADP1 in future iGEM projects. Through our iGEM project, we show that ADP1 is a viable and advantageous chassis organism for synthetic biology and can be used by future iGEM teams for their own creative and scientific endeavors.

Golden Gate Assembly (NEB)

Creating the Exogenous DNA for ADP1 Transformation

General ADP1 Transformation and Genome Manipulation

Overnight Culture

Glycerol Stocks

Polymerase Chain Reaction (PCR)

Gel Electrophoresis

Competent Cells

Spot Plating