Notebook
- Week 1 (June 4-8)
- •Made competent cells
- Week 2 (June 11-15)
- •Testing competent cells using iGem DNA transformation samples.
- • Tested to see if they grew better on a freezer block versus an ice bath.
- • Ran colony PCR and plated bacteria on antimicrobial plate.
- • cPCR, chloramphenicol, and culturing protocols, made agarose gel
- • gel electrophoresis for cPCR, nucleic acid purification plasmid
- Week 3 (June 18-22)
- •Made chloramphenicol LB plates
- •Restriction enzyme digest of iGem plasmid PSB1C3 using EcoRI and PstI
- •Made digest master mix; ran an ezyme digest of iGem plasmids PSB1A3 and PSB1K3 Performed ligation of
- PSB1A3 and PSB1K3; transformed PSB1A3 in competent cells on ampicillin plate
- •Created a 50mg/mL kanamycin solution and then alloquoted it to about 80 1.5mL tubes.
- •Inoculated LB with ampicillin colonies (with insert).
- •Streaked ampicillin (IA3) colony onto new plates.
- •Conducted cPCR for IA3 colonies with insert.
- Week 4 (June 25-29)
- •Conducted cPCR for PSB1K3.
- •Ran 1% agarose gel of PSBIA3 with the insert (colonies that were re-streaked).