Team:Nevada/Notebook/Temp2

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Contents

Week 5 - June 27th- July 3rd

E. Coli

Pyruvate Decarboxylase & Alcohol Dehydrogenase

JC: To insert PDC/ADH genes into the σ70 constitutive promoter plasmid, 0.5ug of σ70/pSB1A3 (2190bp) was digested with SpeI and PstI and 0.5ug of PDC/ADH/pSB1C3 (5124bp) was digested with XbaI and PstI. These digestions cut out the red fluorescent protein (rfp) tag on pSB1A3 (rfp=885bp) to aid in colony selection. Digestions were run on a 1% agarose gel, and bands obtained confirmed PDC/ADH (3054bp), pSB1C3 (2070bp), σ70/pSB1A3 (2190), and rfp (885bp). Incomplete digestion of PDC/ADH/pSB1C3 was apparent for sample #8, so the sample was re-digested before ligation.

PDC/ADH-XbaI/PstI and σ70/pSB1A3-SpeI/PstI were ligated and transformed into NEB10 β cells. Single colonies from LB-Ampicillin plates were selected and tested on LB-Amp and LB-Chlor plates. No usable colonies were obtained (all were light pink and grew on both LB-Chl and LB-Amp plates).=(

Cyano
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Enzymology
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Media
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Week 6 - July 4th-10th

E. Coli
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Cyano
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Enzymology
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Week 7 - July 11th-17th

E. Coli
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Enzymology
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Week 8 - July 18th-24th

E. Coli
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Enzymology
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