EXPLORE

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WHAT ARE WE DOING HERE?

Synthetic Biology is largely restricted to well-funded laboratories at major research universities in high income countries. One significant barrier to entry is the capital cost of instruments. The cloning and assembly of BioBricks, for example, includes the transformation of Escherichia coli, which requires the purchase of a refrigerated centrifuge and an ultra-cold freezer. Here we assemble BioBrick-compatible shuttle vectors for Acinetobacter baylyi ADP1, a naturally competent relative of E. coli that grows as rapidly under identical conditions. We will show that A. baylyi can be transformed with recombinant DNA simply by adding ligation reactions to mid-log cultures; transformants are selected as usual by spreading them onto LB agar plates supplemented with the appropriate antibiotics (kanamycin, spectinomycin, tetracycline, cefotaxime or amikacin). These experiments will show how BioBricks can be constructed and assembled in modestly funded laboratories in community colleges, high schools and even private homes. The resulting plasmid constructs retain their pSB1C3 backbones and will thus remain compatible with the BioBrick standard and capable of replication in the widely used E. coli chassis.

PROJECT

Description

Synthetic Biology is largely restricted to well-funded laboratories at major research universities in high income countries. One significant barrier to entry is the capital cost of instruments. The cloning and assembly of BioBricks, for example, includes the transformation of Escherichia coli, which requires the purchase of a refrigerated centrifuge and an ultra-cold freezer. Here we assemble BioBrick-compatible shuttlevectors for Acinetobacter baylyi ADP1, a naturally competent relative of E. coli that grows as rapidly under identical conditions. We will show that A. baylyi can be transformed with recombinant DNA simply by adding ligation reactions to mid-log cultures; transformants are selected as usual by spreading them onto LB agar plates supplemented with the appropriate antibiotics (kanamycin, spectinomycin, tetracycline, cefotaxime or amikacin). These experiments will show how BioBricks can be constructed and assembled in modestly funded laboratories in community colleges, high schools and even private homes. The resulting plasmid constructs retain their pSB1C3 backbones and will thus remain compatible with the BioBrick standard and capable of replication in the widely used E. coli chassis.

Design, Protocol, and Experiments

Synthetic Biology is largely restricted to well-funded laboratories at major research universities in high income countries. One significant barrier to entry is the capital cost of instruments. The cloning and assembly of BioBricks, for example, includes the transformation of Escherichia coli, which requires the purchase of a refrigerated centrifuge and an ultra-cold freezer. Here we assemble BioBrick-compatible shuttlevectors for Acinetobacter baylyi ADP1, a naturally competent relative of E. coli that grows as rapidly under identical conditions. We will show that A. baylyi can be transformed with recombinant DNA simply by adding ligation reactions to mid-log cultures; transformants are selected as usual by spreading them onto LB agar plates supplemented with the appropriate antibiotics (kanamycin, spectinomycin, tetracycline, cefotaxime or amikacin). These experiments will show how BioBricks can be constructed and assembled in modestly funded laboratories in community colleges, high schools and even private homes. The resulting plasmid constructs retain their pSB1C3 backbones and will thus remain compatible with the BioBrick standard and capable of replication in the widely used E. coli chassis.

Results

Synthetic Biology is largely restricted to well-funded laboratories at major research universities in high income countries. One significant barrier to entry is the capital cost of instruments. The cloning and assembly of BioBricks, for example, includes the transformation of Escherichia coli, which requires the purchase of a refrigerated centrifuge and an ultra-cold freezer. Here we assemble BioBrick-compatible shuttlevectors for Acinetobacter baylyi ADP1, a naturally competent relative of E. coli that grows as rapidly under identical conditions. We will show that A. baylyi can be transformed with recombinant DNA simply by adding ligation reactions to mid-log cultures; transformants are selected as usual by spreading them onto LB agar plates supplemented with the appropriate antibiotics (kanamycin, spectinomycin, tetracycline, cefotaxime or amikacin). These experiments will show how BioBricks can be constructed and assembled in modestly funded laboratories in community colleges, high schools and even private homes. The resulting plasmid constructs retain their pSB1C3 backbones and will thus remain compatible with the BioBrick standard and capable of replication in the widely used E. coli chassis.

NOTEBOOK

We've recorded all of our research achievements and blunders along the way in an online notebook, so you can experience our excitement, thoughts, and the challenges we've encountered and overcome throughout this process.