IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/29
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Results of transformation with our ligation products
Single colony PCR to check our transformation
- Single colony PCR : 13μL of SDW+cells, 5μL of Master Mix, 1μL of VF and 1μL of VR (and plate each single colony) - Load a gel (1.3% agarose) : 5μL of PCR products + 1μL of dye (only 1μL of 100b ladder) In the death plasmid, the VF-VR size is about 280b.
- run again on a e-gel
Stock of ECE112- Bulk up ECE112 (Amp100) rfom 10mL LB (28/08), take 1mL and add to 100mL of LB in big flask - Grow overnight in 37°C incubator with vigorous shaking - Aliquot in 10mL tubes, pellet and freeze cell pellets Transformation of glycerol stock
- TTransfer into Eppendorf tubes and spin down for 30min -Remove glycerol and add medium B - Add ECE112 and ECE153, and one control tube - incubate for 30min in 37°C - Plate out ECE112 (Cm5), ECE153 (Spc50), DNA less (blank, Cm5, Spc50) Plate biobricks from MIT
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