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Week 7

Monday

Wet Work

Amplification

Colony PCR of 8 colonies picked from 391 in pSB3K3 transformants, 8 colonies picked from 374 in pSB3K3 transformants, 8 colonies picked from 384 in pSB3K3, and 1 colony picked from 381 in pSB3K3 transformants confirmed to contain an insert of the right length. Overnight cultures of each of those colonies as well. No 374 constructs were found to contain inserts of the right length, however 4 384 constructs were of the correct length. The 381 construct was confirmed once again to be the correct length.

Overnight,

• Restirction digests of pSB3K3, 380, 382, 385, and 390.
• Colony PCR of 10 new 391 in pSB3K3 transformants, another 374 in pSB3K3 transformants

MelA BioBrick

PCR done of the origonal MelA and the MelA inbtermediate to biobrick (without the second PstI site). This was too create enough DNA for a resitriction digest to be carried out to check that the previous weeks PCRs had been sucessful. After PCR, the products were run on a gel and extracted.

We also tested the Primer C again, as it didn't work last time. Three PCR's were done:

• Primer F and Primer C used last time
• Primer F and a new aliquot of Primer C
• Primer F and revortexed and re-aliquoted primer C from the origonal vial

Dry Work

We found the phzM and phzS pigments in the registry!

• BBa_I723024 (phzM)
• BBa_I723025 (phzS)

This means we can now start working on these pigments, connecting them to processing and logic systems.