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| | <!-- {{#calendar: title=Lethbridge_CCS |year=2008 | month=05}} --> | | <!-- {{#calendar: title=Lethbridge_CCS |year=2008 | month=05}} --> |
| | + | [[Image:Lethbridge_CCS_logo.png|Logo|100px]] [[Team:Lethbridge_CCS|Team Home]] |
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| - | === 2 Sept 2008 === | + | <!-- === July & August === --> |
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| - | (Peter, Paul, Elizabeth, Glenda, Nathan)
| + | '''[[Team:Lethbridge_CCS/Notebook/July & August| July & August Notebook]]''' |
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| - | * Resuspended our primers (psB1A7 forward and reverse; TetR-GFP forward and reverse) to 100 μM.
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| - | * Made 10-fold dilutions of each primer
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| - | * Set up and ran PCR on psB1A7
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| - | ** varied concentration of vector (1 x, 1/10 x, 1/100 x) and annealing temperatures (58°C, 62°C, 66°C), giving nine variations;
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| - | ** used materials and amounts as per Phusion polymerase kit
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| - | - Cycling temperatures and times for PCR
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| - | Initial denaturation 98°C 30 sec ___
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| - | Denaturation 98°C 10 sec |
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| - | Annealing 58/62/66 °C 30 sec |--- 30 cycles
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| - | Extension 72°C 1 min ___|
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| - | Final extension 72°C 10 min
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| - | Hold 4°C hold
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| - | === 3 Sept 2008 ===
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| - | (Peter, Paul, Elizabeth, Marc, Glenda, Nathan)
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| - | * PCR of pSB1A7 from yesterday worked well; approximate size was expected to be ~2500 bp, which was verified (see below)
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| - | ** [[Image:PCRpSB1A7]]
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| - | * did PCR with TetR-GFP using same 9 samples and cycling times/temperatures as yesterday except temperature gradient was 57°C-61°C-65°
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| - | === 6 Sept 2008 ===
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| - | (Peter, Paul, Marc, Glenda, Nathan)
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| - | * looked at PCR results from 3 Sept; some extra bands (see picture)
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| - | ** [[Image:PCRTetR-GFP]]
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| - | * decided to do Agarose gel purification of pSB1A7 from 2 Sept and TetR-GFP from 3 Sept
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| - | * used pooled samples as follows:
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| - | ** pSB1A7: 1/100x at 62°C,1x and 1/10x at 66°C
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| - | ** TetR-GFP: all three 1x samples
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| - | * used Fermentas 1kb GeneRuler; ran at 1000 V for 25 minutes
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| - | * used MinElute Gel Extraction kit to purify
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| | + | '''[[Team:Lethbridge_CCS/Notebook/September|September Notebook]]''' |
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| | + | '''[[Team:Lethbridge_CCS/Notebook/October|October Notebook]]''' |
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| | + | <div style="display:block; text-align:center; border:2px solid #6C5; width:500px; margin:auto; margin-top:3em; padding:5px;"> |
| | + | <p> [[Team:Lethbridge_CCS | Main Page]] | [[Team:Lethbridge_CCS/Team | Team ]] | [[Team:Lethbridge_CCS/Project | Project ]] | [[Team:Lethbridge_CCS/Notebook | Lab Notebook]] </p> |
| | + | </div> |
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| | <!-- ==Notebook== | | <!-- ==Notebook== |
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