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Team:Calgary/23 June 2009
From 2009.igem.org
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| - | + | JUNE 23, 2009 | |
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| - | + | Multiple Miniprep isolations | |
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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 NotI Digest | + | Verification of NotI Digest |
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*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 | *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 | + | *Started with Plasmid Isolation this morning (Sigma). |
*Used 1000 Nanodrop Spectrophotometer to determine concentrations. | *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. | *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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| - | + | 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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| - | + | Verify B0015-R0040 and luxOU-B0015 construction | |
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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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| - | + | Isolating LuxPQ in psB1AK3 from overnights | |
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| - | + | 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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| - | + | 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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Latest revision as of 06:26, 20 October 2009
UNIVERSITY OF CALGARY
