Split IFP & GFP  - Week 1

1        CPXR extraction

Extraction of CPXR from genome of E. Coli strain K-12 MG1655

1.1       Material and methods

Phusion HF PCR Protocol, with corresponding primers (cf primers datasheet). Program PCR: denaturation time of 3 minutes because of the large size of the genome.

1.2       Results

1.2.1     Data

1.2.2     Interpretation

CPXR expected size : 699 bp

Amplification of CPXR worked as expected.

2        Amplification of iGEM plasmid pSB1C3 to insert CPXR

Amplification of pSB1C1 backbone from Cambridge biobrick K325219 (containing Red Firefly Luciferase and LRA under pBAD. This backbone contains the RBS, an arabinose-inducible pBAD promoter, iGEM prefix (containing ECORI and xbaI restriction sites) and suffix (contaning SpeI and PstI restriction sites), a chloramphenicol resistance gene, and an origin of replicaiton.

2.1       Material and methods

Using suitable plasmids, opener1 and opener2 (see primers datasheet), we did a phusion PCR (see protocol).

2.2       Results

2.2.1     Data