Team:Columbia-Cooper-NYC/Columbia notebook 2
From 2012.igem.org
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=== Week 7 === | === Week 7 === | ||
+ | ==== Wednesday, 15th ==== | ||
+ | * Diluted bacteria cultures with following plasmids to 200x LB/CAM and placed in 37°C shaker | ||
+ | *# IPTG-Upps 5-Low Copy (CAM) | ||
+ | *# IPTG-Upps 6-Low Copy (CAM) | ||
+ | *# IPTG-Kill gene-Low Copy (CAM) | ||
==== Thursday, 16th ==== | ==== Thursday, 16th ==== | ||
* Measured OD 600 for 200x diluted bacterial solution containing plasmids with promoter inducible with IPTG | * Measured OD 600 for 200x diluted bacterial solution containing plasmids with promoter inducible with IPTG |
Revision as of 20:31, 16 August 2012
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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
Thursday, 19th
- Purified GFP using standard DNA purification protocol
- Prepared glycerol stock solution (500µl GFP/500µl 80% glycerol) and stored at -80°C
Week 4
Tuesday, 24th
- Re-hydrated four biobrick parts according to standard iGEM re-hydration protocol
- Inducible plasmid (pSB1AK3-J04500) from plate 4, 12A
- GFP (pSB1A2-E0040) from plate 1, 14K
- High copy plasmid pSB1T3-J044500 from plate 1, 7A
- Low copy plasmid (pSB3C5-J044500) from plate 1, 3C
- Electroporated competent E. coli with each of the four above genes separetely
Wednesday, 25th
Thursday, 26th
Friday, 27th
Saturday, 28th
Week 5
Week 6
Week 7
Wednesday, 15th
- Diluted bacteria cultures with following plasmids to 200x LB/CAM and placed in 37°C shaker
- IPTG-Upps 5-Low Copy (CAM)
- IPTG-Upps 6-Low Copy (CAM)
- IPTG-Kill gene-Low Copy (CAM)
Thursday, 16th
- Measured OD 600 for 200x diluted bacterial solution containing plasmids with promoter inducible with IPTG
- IPTG-Upps 5-Low Copy (CAM): .040A
- IPTG-Upps 6-Low Copy (CAM): .032A
- IPTG-Kill gene-Low Copy (CAM): .028A
- Noted that cell concentration was dense, decided to dilute solution with 75µl cells and 925µl LB/CAM solution
- Placed diluted cell solution into 37°C incubator for 40 minutes
- Inserted 1µl of 1M IPTG into cultures
- Placed all samples in 37°C overnight