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From 2014.igem.org

Made electrocompotent MACH1 and PHL cells

We ran the pbBE1a backbone from yesterday on a gel and this time we got bands

We gel purified the backbone and hybB insert PCR products for Gibson→ Ran Gibson Rxn → Transformed Gibson Rxn into both MACH1 and Turbo cells→ Plated

PCR (Red pigment, BLux, RLux, HSP, asr) for Gibson reaction.

Sequencing: Chromoproteins (pHBD32a, 32b, 33a, 33b, 34a, 34b) with Fwd prefix HBD31 and Rev suffix HBD34 pSB3K3 (pHBD35a, 35b) with fwd&rev suffix (HBD33 & HBD34) and fwd2&rev2 noncoding prefix (HBD29 & HBD30) We received the sequencing primers for internally sequencing the red pigment however we need more of HBD4 (Red pigment) so we need to wait for the miniprep tomorrow.

“To Do” -Design internal lux sequencing primers -Order UV promoter/Orange pigment biobricks when we go to pick up the rest of our chromoprotein orders from the igem hq tomorrow -Design gibson primers for oxygen promoters -Make more Cm and Carb plates