From 2012.igem.org
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| | After consultation with Hila, I decided to digest the plasmid (the original one, not the PCR-amplified) with Pst1 and then run it on a gel, so it will run in his linear form- The results wasn't comforting- there wasn't any band! I need to start all over again tomorrow... | | After consultation with Hila, I decided to digest the plasmid (the original one, not the PCR-amplified) with Pst1 and then run it on a gel, so it will run in his linear form- The results wasn't comforting- there wasn't any band! I need to start all over again tomorrow... |
| | ==Asaf== | | ==Asaf== |
| | + | I ran the PCR products of the Riboswitch on extraction gel, and cut the bands from the gel. |
| | | | |
| | + | ==Asaf and Shahar == |
| | + | PCR for extracting the different polymerase genes from the different plasmids. |
| | + | |
| | ==Hila== | | ==Hila== |
| | - ligation for the MCS insert to the cut pSB1C3 plasmid. <br> | | - ligation for the MCS insert to the cut pSB1C3 plasmid. <br> |
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| | <br> -Transformation of the ligated plasmid into TOP10 competent strain. | | <br> -Transformation of the ligated plasmid into TOP10 competent strain. |
| | <br> -Plating ligated plasmid+control on Kanamycin plates and growing at 37C overnight. | | <br> -Plating ligated plasmid+control on Kanamycin plates and growing at 37C overnight. |
| - | ==Shahar==
| + | |
| | | | |
| | ==Rachel== | | ==Rachel== |
| | | | |
| | }} | | }} |
Revision as of 11:33, 30 August 2012
Team:Technion/22_August_2012
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_March_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
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| [http://2012.igem.org/Team:Technion/25_April_2012 25]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_April_2012&preload=Team:Technion/Template:NewDate&action=edit 27]
| [http://2012.igem.org/Team:Technion/28_April_2012 28]
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| [http://2012.igem.org/Team:Technion/1_May_2012 1]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_May_2012&preload=Team:Technion/Template:NewDate&action=edit 3]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_May_2012&preload=Team:Technion/Template:NewDate&action=edit 4]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_May_2012&preload=Team:Technion/Template:NewDate&action=edit 5]
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| [http://2012.igem.org/Team:Technion/10_May_2012 10]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_May_2012&preload=Team:Technion/Template:NewDate&action=edit 15]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_May_2012&preload=Team:Technion/Template:NewDate&action=edit 16]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_May_2012&preload=Team:Technion/Template:NewDate&action=edit 17]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_May_2012&preload=Team:Technion/Template:NewDate&action=edit 26]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_May_2012&preload=Team:Technion/Template:NewDate&action=edit 30]
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| [http://2012.igem.org/Team:Technion/3_June_2012 3]
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| [http://2012.igem.org/Team:Technion/5_June_2012 5]
| [http://2012.igem.org/Team:Technion/6_June_2012 6]
| [http://2012.igem.org/Team:Technion/7_June_2012 7]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_June_2012&preload=Team:Technion/Template:NewDate&action=edit 8]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_June_2012&preload=Team:Technion/Template:NewDate&action=edit 9]
| [http://2012.igem.org/Team:Technion/10_June_2012 10]
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| [http://2012.igem.org/Team:Technion/14_June_2012 14]
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| [http://2012.igem.org/Team:Technion/7_August_2012 7]
| [http://2012.igem.org/Team:Technion/8_August_2012 8]
| [http://2012.igem.org/Team:Technion/9_August_2012 9]
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| [http://2012.igem.org/Team:Technion/12_August_2012 12]
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| [http://2012.igem.org/Team:Technion/14_August_2012 14]
| [http://2012.igem.org/Team:Technion/15_August_2012 15]
| [http://2012.igem.org/Team:Technion/16_August_2012 16]
| [http://2012.igem.org/Team:Technion/17_August_2012 17]
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| [http://2012.igem.org/Team:Technion/19_August_2012 19]
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| [http://2012.igem.org/Team:Technion/21_August_2012 21]
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| [http://2012.igem.org/Team:Technion/28_August_2012 28]
| [http://2012.igem.org/Team:Technion/29_August_2012 29]
| [http://2012.igem.org/Team:Technion/30_August_2012 30]
| [http://2012.igem.org/Team:Technion/31_August_2012 31]
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| September
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| M | T | W | T | F | S | S
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| [http://2012.igem.org/Team:Technion/1_September_2012 1]
| [http://2012.igem.org/Team:Technion/2_September_2012 2]
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| [http://2012.igem.org/Team:Technion/3_September_2012 3]
| [http://2012.igem.org/Team:Technion/4_September_2012 4]
| [http://2012.igem.org/Team:Technion/5_September_2012 5]
| [http://2012.igem.org/Team:Technion/6_September_2012 6]
| [http://2012.igem.org/Team:Technion/7_September_2012 7]
| [http://2012.igem.org/Team:Technion/8_September_2012 8]
| [http://2012.igem.org/Team:Technion/9_September_2012 9]
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| [http://2012.igem.org/Team:Technion/10_September_2012 10]
| [http://2012.igem.org/Team:Technion/11_September_2012 11]
| [http://2012.igem.org/Team:Technion/12_September_2012 12]
| [http://2012.igem.org/Team:Technion/13_September_2012 13]
| [http://2012.igem.org/Team:Technion/14_September_2012 14]
| [http://2012.igem.org/Team:Technion/15_September_2012 15]
| [http://2012.igem.org/Team:Technion/16_September_2012 16]
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| [http://2012.igem.org/Team:Technion/17_September_2012 17]
| [http://2012.igem.org/Team:Technion/18_September_2012 18]
| [http://2012.igem.org/Team:Technion/19_September_2012 19]
| [http://2012.igem.org/Team:Technion/20_September_2012 20]
| [http://2012.igem.org/Team:Technion/21_September_2012 21]
| [http://2012.igem.org/Team:Technion/22_September_2012 22]
| [http://2012.igem.org/Team:Technion/23_September_2012 23]
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| [http://2012.igem.org/Team:Technion/24_September_2012 24]
| [http://2012.igem.org/Team:Technion/25_September_2012 25]
| [http://2012.igem.org/Team:Technion/26_September_2012 26]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_September_2012&preload=Team:Technion/Template:NewDate&action=edit 27]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_September_2012&preload=Team:Technion/Template:NewDate&action=edit 28]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_September_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_September_2012&preload=Team:Technion/Template:NewDate&action=edit 30]
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| October
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| M | T | W | T | F | S | S
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_October_2012&preload=Team:Technion/Template:NewDate&action=edit 1]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_October_2012&preload=Team:Technion/Template:NewDate&action=edit 2]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_October_2012&preload=Team:Technion/Template:NewDate&action=edit 3]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_October_2012&preload=Team:Technion/Template:NewDate&action=edit 4]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_October_2012&preload=Team:Technion/Template:NewDate&action=edit 5]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_October_2012&preload=Team:Technion/Template:NewDate&action=edit 6]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_October_2012&preload=Team:Technion/Template:NewDate&action=edit 7]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_October_2012&preload=Team:Technion/Template:NewDate&action=edit 8]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_October_2012&preload=Team:Technion/Template:NewDate&action=edit 9]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_October_2012&preload=Team:Technion/Template:NewDate&action=edit 10]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_October_2012&preload=Team:Technion/Template:NewDate&action=edit 11]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_October_2012&preload=Team:Technion/Template:NewDate&action=edit 12]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_October_2012&preload=Team:Technion/Template:NewDate&action=edit 13]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_October_2012&preload=Team:Technion/Template:NewDate&action=edit 14]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_October_2012&preload=Team:Technion/Template:NewDate&action=edit 15]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_October_2012&preload=Team:Technion/Template:NewDate&action=edit 16]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_October_2012&preload=Team:Technion/Template:NewDate&action=edit 17]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_October_2012&preload=Team:Technion/Template:NewDate&action=edit 18]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_October_2012&preload=Team:Technion/Template:NewDate&action=edit 19]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_October_2012&preload=Team:Technion/Template:NewDate&action=edit 20]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_October_2012&preload=Team:Technion/Template:NewDate&action=edit 21]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_October_2012&preload=Team:Technion/Template:NewDate&action=edit 22]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_October_2012&preload=Team:Technion/Template:NewDate&action=edit 23]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_October_2012&preload=Team:Technion/Template:NewDate&action=edit 24]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_October_2012&preload=Team:Technion/Template:NewDate&action=edit 25]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_October_2012&preload=Team:Technion/Template:NewDate&action=edit 26]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_October_2012&preload=Team:Technion/Template:NewDate&action=edit 27]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_October_2012&preload=Team:Technion/Template:NewDate&action=edit 28]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_October_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_October_2012&preload=Team:Technion/Template:NewDate&action=edit 30]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/31_October_2012&preload=Team:Technion/Template:NewDate&action=edit 31]
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| November
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_November_2012&preload=Team:Technion/Template:NewDate&action=edit 1]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_November_2012&preload=Team:Technion/Template:NewDate&action=edit 2]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_November_2012&preload=Team:Technion/Template:NewDate&action=edit 3]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_November_2012&preload=Team:Technion/Template:NewDate&action=edit 4]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_November_2012&preload=Team:Technion/Template:NewDate&action=edit 5]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_November_2012&preload=Team:Technion/Template:NewDate&action=edit 6]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_November_2012&preload=Team:Technion/Template:NewDate&action=edit 7]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_November_2012&preload=Team:Technion/Template:NewDate&action=edit 8]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_November_2012&preload=Team:Technion/Template:NewDate&action=edit 9]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_November_2012&preload=Team:Technion/Template:NewDate&action=edit 10]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_November_2012&preload=Team:Technion/Template:NewDate&action=edit 11]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_November_2012&preload=Team:Technion/Template:NewDate&action=edit 12]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_November_2012&preload=Team:Technion/Template:NewDate&action=edit 13]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_November_2012&preload=Team:Technion/Template:NewDate&action=edit 14]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_November_2012&preload=Team:Technion/Template:NewDate&action=edit 15]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_November_2012&preload=Team:Technion/Template:NewDate&action=edit 16]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_November_2012&preload=Team:Technion/Template:NewDate&action=edit 17]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_November_2012&preload=Team:Technion/Template:NewDate&action=edit 18]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_November_2012&preload=Team:Technion/Template:NewDate&action=edit 19]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_November_2012&preload=Team:Technion/Template:NewDate&action=edit 20]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_November_2012&preload=Team:Technion/Template:NewDate&action=edit 21]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_November_2012&preload=Team:Technion/Template:NewDate&action=edit 22]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_November_2012&preload=Team:Technion/Template:NewDate&action=edit 23]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_November_2012&preload=Team:Technion/Template:NewDate&action=edit 24]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_November_2012&preload=Team:Technion/Template:NewDate&action=edit 25]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_November_2012&preload=Team:Technion/Template:NewDate&action=edit 26]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_November_2012&preload=Team:Technion/Template:NewDate&action=edit 27]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_November_2012&preload=Team:Technion/Template:NewDate&action=edit 28]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_November_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_November_2012&preload=Team:Technion/Template:NewDate&action=edit 30]
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Ilya
- Transformation of R0062+Fus1del in pSB1C3 and in pSB3C5 and of Fus2del in pSB1C3 and in pSB3C5. These clones will serve as positive controls for the plate reader assay. I think correct clones should give red colonies tomorrow.
Inbal
- 2 starters of the TOP10 bacteria containing the SP6 gene in pSB1C3 plasmid, I grow them for 8 hours in 37C incubator, then I mini preped the plasmidial DNA.
Also I did deletion PCR for the Bba_I13522 (the conc. of the template: 105ng/ul) , excluding the terminators in the part, I ran the PCR products on 1% agarose gel for 40 minutes. The expected band is 3kb size approximately. The results: A 4000bp band size (likely there was a mistake in the experiment).
After consultation with Hila, I decided to digest the plasmid (the original one, not the PCR-amplified) with Pst1 and then run it on a gel, so it will run in his linear form- The results wasn't comforting- there wasn't any band! I need to start all over again tomorrow...
Asaf
I ran the PCR products of the Riboswitch on extraction gel, and cut the bands from the gel.
Asaf and Shahar
PCR for extracting the different polymerase genes from the different plasmids.
Hila
- ligation for the MCS insert to the cut pSB1C3 plasmid.
- chemical transformation of the ligated product to Top 10 - Rb competent bacteria.
- recovery for pSB1C3+fuz_1 bacteria from glycerol stock.
please cross your fingers…
Lior
Great Success!! We got colonies! more ALP than xyIE.
We tried antother run for Chris's plasmids- this time with Tm of 62, 7 degrees more than last time.
And it worked! We got 4 chubby bands.
Colony PCR for ALP in order to verify the insert within the pT7 plasmid, we got 4 colonies out of 5 (4/5)
Noa
Evgeni
-Ligation of pPROLar cut with BamHI and HindIII with Bba_015. Reaction with no insert serves as control.
-Transformation of the ligated plasmid into TOP10 competent strain.
-Plating ligated plasmid+control on Kanamycin plates and growing at 37C overnight.
Rachel
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