Team:Columbia-Cooper-NYC/Columbia notebook 2
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Columbia Genetics Lab Notebook
July, 2012
Week 1
Thursday, 5th
- Re-hydrated plasmids with 50µl of LB and Kanamycin solution
- Stored solution at 37°C incubator overnight
Friday, 6th
- Purified pET26b vector using standard DNA purification protocol
Week 2
Monday, 9th
- Received kill gene Bba-K124017 from plate 3, 20M
- Re-hydrated DNA according to standard iGEM re-hydration protocol
Tuesday, 10th
- Contacted professors at Germany in hopes to receive copies of fungal phytochrome FphA
Wednesday, 11th
- Received confirmation by professors at Germany for FphA to be sent to Columbia University
- Conducted transformation using electroporation with competent bacteria (marked by resistance to Kanamycin)
- Control: 1µl of deionized water with abt. and 60µl of bacteria cells
- Variable: 1µl of re-hydrated kill gene and 60µl of bacteria cells
- Placed both samples after electroporation into 200µl of preprepared LB
- Placed samples in shaker 37°C for 30 minutes
Thursday, 12th
- Grew 1 colony of transformed bacteria in 5mL of Kanamycin and LB solution
- Note: Using pET20b vector over pET26b vector from glycerol stock solution
Friday, 13th
- Isolated 4 samples of plasmid using standard plasmid isolation protocol
- 2 samples: kill gene
- 2 samples: pET20b vector
Week 3
Monday, 16th
- Re-hydrated two biobrick parts in plasmid pSB2K3 according to standard iGEM re-hydration protocol
- BBa-I16009 (PcyA) from plate 1, 20F
- BBa-I16008 (ho1) from plate 2, 13J
- Electroporated 1µl of each biobrick into separate E. coli at 1800V
- Added 100µl LB broth into each sample
- Placed samples at 33.4°C for 20 minutes
- Samples were plated to be grown overnight
Tuesday, 17th
- Placed 5ml each of LB/Kan into two centrifuge tube for PCB creation
- Label P: PcyA
- Label h: ho1
- Placed samples in 37°C incubator
Wednesday, 18th
- Purified ho1 and PcyA plasmids using standard DNA purification protocol
- Placed purified DNA into glycerol stock (LB/Kan) and stored at -80°C