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| - | June 23, 2009
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| - | Descriptive Title of What You’re Doing
| + | Multiple Miniprep isolations |
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| - | WIKI CODING HERE
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| - | CHINMOYEE
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| | + | * Attempted 15 miniprep reactions on both non-mutated and mutated <i>luxCDABE</i> plasmid. |
| | + | * The isolated plasmid was pooled together and attempted to concentrate the plasmid by spinning the tube in a vacifuge. After several hours (2.5 hours) the concentration was higher. |
| | + | * Tried ethanol precipitation earlier in the day but there was no precipitate found when done. |
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| - | Verification Colony PCR of BBK LuxOD47E in psBA1AC3 Vector | + | Verification of NotI Digest |
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| - | *Performed colony PCR again with the proper PCR Buffer this time. PCR products were visualized on a 1% agarose gel run at 120 V with Generuler 1.0 KB+ DNA Ladder. Lanes 1-7 are LuxOD47E in psB1AC3 cut with XbaI / PstI. Lane 8 is a negative control with only Mastermix + ddH2O. | + | *Objective: To do one last verification that LuxOD47E is in the psB1AC3 vector before we send it down for DNA Sequecning. We will do a NotI digest with |
| - | *See gel photo below. | + | *Started with Plasmid Isolation this morning (Sigma). |
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| + | *Used 1000 Nanodrop Spectrophotometer to determine concentrations. |
| | + | *Set up a restriction digest with NotI enzyme and REact Buffer 3, left to digest in the 37 C waterbath overnight. |
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| + | Marketing for 23rd June 2009 |
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| - | WIKI CODING HERE
| + | Today, I helped finish the July newsletter which will be ready be next week to sent out to our sponsors. Also I e-mailed some research and development companies and will do a follow-up soon. |
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| - | WIKI CODING HERE
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| - | Descriptive Title of What You're Doing
| + | Verify B0015-R0040 and luxOU-B0015 construction |
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| - | WIKI CODING HERE
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| | + | [[Image:Calgary_luxOU_B0015-R0040+luxOU-B0015_NotI.jpg | 500px]] |
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| | + | Figure. 1% agarose gel (100V) for NotI verification digest of construction yielding BBa_B0015-BBa_R0040 and luxOU(XbaI or EcoRI)-BBa_B0015. 200ng of plasmid was cut with NotI for 2 hours at 37oC. BBa_B0015-BBa_R0040 C1 is lane 2) and C4 is lane 4. luxOU(XbaI)-BBa_B0015 C3 is in lane 4 and C4 is in lane 5. luxOU(EcoRI)-BBa_B0015 C3 is in lane 6 and C4 in lane 7. Uncut luxOU in psB1AC3 (EcoRI C1 and XbaI C2) were used as positive controls in lanes 8 and 9, respectively. Expected band sizes for the BBa_B0015-BBa_R0040 construction are 3.2kb (psB1AK3) and 183bp. Expected band sizes for the luxOU-BBa_B0015 construction are 3kb (psB1AC3) and 2.1kb. 5μL of GeneRuler 1kb Plus DNA Ladder was loaded. |
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| | + | BBa_B0015-BBa_R0040 C1 and luxOU(XbaI)-BBa_B0015 C4 were sent for sequencing with BBK-CP-F and BBK-CP-R as sequencing primers. Sequencing results were checked for vector contamination using “VecScreen”, protein sequences using “Blastx” and sequence conformation using “Blast2” (National Center for Biotechnology Information, MD). |
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| - | Descriptive Title of What You're Doing
| + | Isolating LuxPQ in psB1AK3 from overnights |
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| - | WIKI CODING HERE
| + | Purpose: isolate LuxPQ in psB1AK3 from overnights of Colony 4 to continue construction of signaling circuit. Plasmid was isolated from colony 4 of luxPQ. This was done using the SIGMA’s Genelute plasmid Miniprep Kit. Plasmid purity and concentration were measured using the NanoDrop Spectrophotometer. |
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| | To obtain more isolated R0040, B0015, J1302, and Pqrr4, miniprep (plasmid isolation) was done on them. | | To obtain more isolated R0040, B0015, J1302, and Pqrr4, miniprep (plasmid isolation) was done on them. |
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| + | Blog Making |
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| - | WIKI CODING HERE
| + | Put together a workable blog and other online accounts for our team to update each other on what they've been up to, with an established schedule and everything :) Now I just have to figure out how to get it onto our wiki, or if I have to transfer each post manually from http://igemcalgary.blogspot.com |
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CAROL
Multiple Miniprep isolations
- Attempted 15 miniprep reactions on both non-mutated and mutated luxCDABE plasmid.
- The isolated plasmid was pooled together and attempted to concentrate the plasmid by spinning the tube in a vacifuge. After several hours (2.5 hours) the concentration was higher.
- Tried ethanol precipitation earlier in the day but there was no precipitate found when done.
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EMILY
Verification of NotI Digest
- Objective: To do one last verification that LuxOD47E is in the psB1AC3 vector before we send it down for DNA Sequecning. We will do a NotI digest with
- Started with Plasmid Isolation this morning (Sigma).
- Used 1000 Nanodrop Spectrophotometer to determine concentrations.
- Set up a restriction digest with NotI enzyme and REact Buffer 3, left to digest in the 37 C waterbath overnight.
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FAHD
Marketing for 23rd June 2009
Today, I helped finish the July newsletter which will be ready be next week to sent out to our sponsors. Also I e-mailed some research and development companies and will do a follow-up soon.
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JAMIE
Verify B0015-R0040 and luxOU-B0015 construction
Figure. 1% agarose gel (100V) for NotI verification digest of construction yielding BBa_B0015-BBa_R0040 and luxOU(XbaI or EcoRI)-BBa_B0015. 200ng of plasmid was cut with NotI for 2 hours at 37oC. BBa_B0015-BBa_R0040 C1 is lane 2) and C4 is lane 4. luxOU(XbaI)-BBa_B0015 C3 is in lane 4 and C4 is in lane 5. luxOU(EcoRI)-BBa_B0015 C3 is in lane 6 and C4 in lane 7. Uncut luxOU in psB1AC3 (EcoRI C1 and XbaI C2) were used as positive controls in lanes 8 and 9, respectively. Expected band sizes for the BBa_B0015-BBa_R0040 construction are 3.2kb (psB1AK3) and 183bp. Expected band sizes for the luxOU-BBa_B0015 construction are 3kb (psB1AC3) and 2.1kb. 5μL of GeneRuler 1kb Plus DNA Ladder was loaded.
BBa_B0015-BBa_R0040 C1 and luxOU(XbaI)-BBa_B0015 C4 were sent for sequencing with BBK-CP-F and BBK-CP-R as sequencing primers. Sequencing results were checked for vector contamination using “VecScreen”, protein sequences using “Blastx” and sequence conformation using “Blast2” (National Center for Biotechnology Information, MD).
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JEREMY
Isolating LuxPQ in psB1AK3 from overnights
Purpose: isolate LuxPQ in psB1AK3 from overnights of Colony 4 to continue construction of signaling circuit. Plasmid was isolated from colony 4 of luxPQ. This was done using the SIGMA’s Genelute plasmid Miniprep Kit. Plasmid purity and concentration were measured using the NanoDrop Spectrophotometer.
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KEVIN
1. Glycerol Stock
Glycerol stocks of R0040, B0015 J13002, and Pqrr4 were made in order to preserve them long term. The addition of glycerl is important because otherwise the cellular membrane would shear.
2. Plasmid Isolation
To obtain more isolated R0040, B0015, J1302, and Pqrr4, miniprep (plasmid isolation) was done on them.
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MANDY
Blog Making
Put together a workable blog and other online accounts for our team to update each other on what they've been up to, with an established schedule and everything :) Now I just have to figure out how to get it onto our wiki, or if I have to transfer each post manually from http://igemcalgary.blogspot.com
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PRIMA
Marketing & Sponsorship
After constantly following up with companies over the past few weeks, I successfully recruited Alberta Research Council to sponsor the University of Calgary's iGEM team. They have generously donated %250 dollars this year.
We also edited the June newsletter and discussed it with the rest of the marketing team prior to sending it to all the companies we had contacted (including those who have declined our proposal).
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VICKI
Sequencing of what I hope will be LuxOD47A BBK
I prepared a sample of the recently-formed-and-colony-PCR'd-and-NotI-restriction-digested LuxOD47A BBk for sequencing. This was done with BBk sequencing primers, in accordance with the sequencing protocol.
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