From 2012.igem.org
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| | __NOTOC__ | | __NOTOC__ |
| | ==Ilya== | | ==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. |
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| | ==Inbal== | | ==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. <br> |
| | + | 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).<br> |
| | + | 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.<br> |
| | + | I also purified the gel extracts. |
| | | | |
| | + | ==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> |
| | - chemical transformation of the ligated product to Top 10 - Rb competent bacteria. <br> | | - chemical transformation of the ligated product to Top 10 - Rb competent bacteria. <br> |
| | + | - recovery for pSB1C3+fuz_1 bacteria from glycerol stock. <br> |
| | + | <br> |
| | + | please cross your fingers… <br> |
| | + | |
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| | ==Lior== | | ==Lior== |
| | + | Great Success!! We got colonies! more ALP than xyIE. <br> |
| | + | We tried antother run for Chris's plasmids- this time with Tm of 62, 7 degrees more than last time. <br> |
| | + | And it worked! We got 4 chubby bands. <br> |
| | + | Colony PCR for ALP in order to verify the insert within the pT7 plasmid, we got 4 colonies out of 5 (4/5) <br> |
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| | ==Noa== | | ==Noa== |
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| | ==Evgeni== | | ==Evgeni== |
| | + | -Ligation of pPROLar cut with BamHI and HindIII with Bba_015. Reaction with no insert serves as control. |
| | + | <br> -Transformation of the ligated plasmid into TOP10 competent strain. |
| | + | <br> -Plating ligated plasmid+control on Kanamycin plates and growing at 37C overnight. |
| | | | |
| - | ==Shahar==
| |
| | | | |
| | ==Rachel== | | ==Rachel== |
| | | | |
| | }} | | }} |
Latest revision as of 11:35, 30 August 2012
Team:Technion/22_August_2012
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| [http://2012.igem.org/Team:Technion/25_April_2012 25]
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| [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/wiki/index.php?title=Team:Technion/9_May_2012&preload=Team:Technion/Template:NewDate&action=edit 9]
| [http://2012.igem.org/Team:Technion/10_May_2012 10]
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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/29_May_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
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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/12_June_2012 12]
| [http://2012.igem.org/Team:Technion/13_June_2012 13]
| [http://2012.igem.org/Team:Technion/14_June_2012 14]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_June_2012&preload=Team:Technion/Template:NewDate&action=edit 28]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_June_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
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| [http://2012.igem.org/Team:Technion/3_July_2012 3]
| [http://2012.igem.org/Team:Technion/4_July_2012 4]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_July_2012&preload=Team:Technion/Template:NewDate&action=edit 5]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_July_2012&preload=Team:Technion/Template:NewDate&action=edit 6]
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| [http://2012.igem.org/Team:Technion/10_July_2012 10]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_July_2012&preload=Team:Technion/Template:NewDate&action=edit 13]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_July_2012&preload=Team:Technion/Template:NewDate&action=edit 14]
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| [http://2012.igem.org/Team:Technion/18_July_2012 18]
| [http://2012.igem.org/Team:Technion/19_July_2012 19]
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| [http://2012.igem.org/Team:Technion/26_July_2012 26]
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| [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_August_2012&preload=Team:Technion/Template:NewDate&action=edit 3]
| [http://2012.igem.org/Team:Technion/4_August_2012 4]
| [http://2012.igem.org/Team:Technion/5_August_2012 5]
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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]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_August_2012&preload=Team:Technion/Template:NewDate&action=edit 10]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_August_2012&preload=Team:Technion/Template:NewDate&action=edit 11]
| [http://2012.igem.org/Team:Technion/12_August_2012 12]
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| [http://2012.igem.org/Team:Technion/13_August_2012 13]
| [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]
| [http://2012.igem.org/Team:Technion/18_August_2012 18]
| [http://2012.igem.org/Team:Technion/19_August_2012 19]
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| [http://2012.igem.org/Team:Technion/20_August_2012 20]
| [http://2012.igem.org/Team:Technion/21_August_2012 21]
| [http://2012.igem.org/Team:Technion/22_August_2012 22]
| [http://2012.igem.org/Team:Technion/23_August_2012 23]
| [http://2012.igem.org/Team:Technion/24_August_2012 24]
| [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_August_2012&preload=Team:Technion/Template:NewDate&action=edit 25]
| [http://2012.igem.org/Team:Technion/26_August_2012 26]
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| [http://2012.igem.org/Team:Technion/27_August_2012 27]
| [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.
I also purified the gel extracts.
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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