From 2012.igem.org
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| | ==Inbal== | | ==Inbal== |
| - | | + | - Restriction for BBa_B0015 (ter+pSB1AK3) with XbaI (3189bp). <br> |
| | + | - 2 starters of T3 RNAP with CM resistance (BBa_K346000). <br> |
| | ==Asaf== | | ==Asaf== |
| | I figured that I don't have enough insert (Riboswitch with polymerase) for ligation with the Psb1c3.<br> | | I figured that I don't have enough insert (Riboswitch with polymerase) for ligation with the Psb1c3.<br> |
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| | I also helped Evgeni and made glycerol stocks of his starters,<br> and also centerfuge the starters so that he could miniprep them tomorrow. | | I also helped Evgeni and made glycerol stocks of his starters,<br> and also centerfuge the starters so that he could miniprep them tomorrow. |
| | ==Hila== | | ==Hila== |
| | + | - Last and final colony PCR for fragments 1C, 4C, 6C and 7C in the pSB1C3_MCS plasmid. No positive colonies were found. <br> |
| | + | - In addition I made a PCR reaction for fragments 1C, 4C, 6C, 7C just in case I won't find positive colonies. The PCR for fragments 1C and 6C didn't work again… <br> |
| | + | - Additional amplification to F4, F4* and F5. In addition today the 250bp overlap primer arrived, so I amplified fragment F4' which will help us in the Gibson assembly calibration. <br> |
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| | ==Lior== | | ==Lior== |
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| | ==Shahar== | | ==Shahar== |
| | + | colony PCR for fragments 1C, 4C, 6C and 7C in the pSB1C3_MCS plasmid. No positive colonies were found. <br> |
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| | ==Rachel== | | ==Rachel== |
| - | | + | -Miniprep for pSB1AK3(BBa_B0015 biobrick) , the concentration was: 262.5 ng/ul.<br> |
| | + | -Digestion of pSB1AK3(BBa_B0015 biobrick) with XbaI, for 2 hours.<br> |
| | + | -Clean the SB1AK3(BBa_B0015) by column cleaning kit.<br> |
| | + | -I ran on gel the restriction products of SB1AK3(BBa_B0015) for 30 minutes, I also ran uncut plasmid as contro. The expected cut plasmid (linear) was 3300bp size. The uncut plasmid (pSB1AK3) ran up to 2200bp.<br> |
| | + | -The plasmid was treated with cip enzyme for 1 hour at 37C. Then it was cleaned bu column cleaning kit, the concentration was 62ng/ul.<br> |
| | + | -4 ligatins: T7*(K1F) RNAP + pSB1AK3, T7*(N4)RNAP + pSB1AK3, T7*(T3)RNAP + pSB1AK3 and control (pSB1AK3 without any insert). The ligation was at 16C overnight. |
| | }} | | }} |
Latest revision as of 11:53, 26 September 2012
Team:Technion/13_September_2012
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| [http://2012.igem.org/Team:Technion/5_June_2012 5]
| [http://2012.igem.org/Team:Technion/6_June_2012 6]
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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]
|
| [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]
|
| [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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| M | T | W | T | F | S | S
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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
- Began working on adding our parts to the registry.
Inbal
- Restriction for BBa_B0015 (ter+pSB1AK3) with XbaI (3189bp).
- 2 starters of T3 RNAP with CM resistance (BBa_K346000).
Asaf
I figured that I don't have enough insert (Riboswitch with polymerase) for ligation with the Psb1c3.
I have decided to extract more insert from gel from the product I got from the fusion PCR with Tm of 55C.
I couldn't find the T7* product. I have already used both products of T7.
So I ran the Riboswitch with the K1F, T3 and N4 polymerase genes on an extraction gel.
I got the expected bands,but I didn't get the 1Kb pattern, and cut the 3.2 kb bands (as I expected from previuos gels).
I cleaned the DNA from the gel.
I cut all of my inserts (Riboswitch with different polymerases) with EcoRI and PstI.
I also helped Evgeni and made glycerol stocks of his starters,
and also centerfuge the starters so that he could miniprep them tomorrow.
Hila
- Last and final colony PCR for fragments 1C, 4C, 6C and 7C in the pSB1C3_MCS plasmid. No positive colonies were found.
- In addition I made a PCR reaction for fragments 1C, 4C, 6C, 7C just in case I won't find positive colonies. The PCR for fragments 1C and 6C didn't work again…
- Additional amplification to F4, F4* and F5. In addition today the 250bp overlap primer arrived, so I amplified fragment F4' which will help us in the Gibson assembly calibration.
Lior
Noa
Evgeni
Shahar
colony PCR for fragments 1C, 4C, 6C and 7C in the pSB1C3_MCS plasmid. No positive colonies were found.
Rachel
-Miniprep for pSB1AK3(BBa_B0015 biobrick) , the concentration was: 262.5 ng/ul.
-Digestion of pSB1AK3(BBa_B0015 biobrick) with XbaI, for 2 hours.
-Clean the SB1AK3(BBa_B0015) by column cleaning kit.
-I ran on gel the restriction products of SB1AK3(BBa_B0015) for 30 minutes, I also ran uncut plasmid as contro. The expected cut plasmid (linear) was 3300bp size. The uncut plasmid (pSB1AK3) ran up to 2200bp.
-The plasmid was treated with cip enzyme for 1 hour at 37C. Then it was cleaned bu column cleaning kit, the concentration was 62ng/ul.
-4 ligatins: T7*(K1F) RNAP + pSB1AK3, T7*(N4)RNAP + pSB1AK3, T7*(T3)RNAP + pSB1AK3 and control (pSB1AK3 without any insert). The ligation was at 16C overnight.
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