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From 2011.igem.org

Contents 1 Thuesday 1.1 Lab Work 1.1.1 A1 1.1.2 A2 1.1.3 B1 1.1.4 Not Plant Cyps 1.1.5 Human CYPs

Thuesday

Ups. We forgot to check the promoter on the biobrick J04450. It's IPTG inducuble. Therefore we had no red colonies. And we were so looking forward to them. But there were colonies and we'll try to give one of them IPTG and see what happens. A2 and B2 ligations didn't work as hoped. (we did'nt have any transformants) So we're trying to different ligation approaches. We let the plasmid and the cyp stand for 1 hour to ligate and we have another edition of cyp and plasmid that will ligate over night in the fridge.

Lab Work

A1

Extended our CYP A1 with prefix and suffix by a PCR approach.

Messed up the purification step - ups!

A2

• Ligation of the BioBrick (CYP79A2) and the plasmid backbone (pSB1C3).
• Transformation of the constructed plasmid into competent cells.

B1

• Ligation of the BioBrick (CYP79 B1) and the plasmid backbone (pSB1C3).
• Transformation of the constructed plasmid into competent cells.

Not Plant Cyps

• Digested the standard plasmid from iGEM pSB1C3 with EcoR1 and Pst1. Note: Make more than you need and keep the lid firmly closed.
• Transformation with pSB1C3 from the iGEM kit on plates with IPTG.
• Transformation of the pSB1C3 transformed XL1-Blue from yesterday on plates with IPTG.

Human CYPs

• Waiting for sequences to return

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