From 2012.igem.org
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| | ==Inbal== | | ==Inbal== |
| - | -I did a research on the SP6 gene that was sent to us from china- I found out the sequence of the plasmid it came with, I also found the map+restriction sites of the plasmid (pACSP6R, or his other commercial name: pACYC184). <br> | + | -I did a research on the SP6 gene that was sent to us from china- I found the sequence of the plasmid it came with, I also found the map+restriction sites of the plasmid (pACSP6R, or his other commercial name: pACYC184). <br> |
| | after that, I digested this plasmid with BamHI+HindIII and ran on 1% agarose gel. The expected bands were: 5600 and 350 bp approximately. The actual bands that I got were 3 light bands: 3kb, 2.5kb and 1.5kb and very light smear above them. Tomorrow I need to send for sequencing this plasmid with 2 primers (that I already have) to verify once and for good the sequence! I hope for the BEST!!!<br> | | after that, I digested this plasmid with BamHI+HindIII and ran on 1% agarose gel. The expected bands were: 5600 and 350 bp approximately. The actual bands that I got were 3 light bands: 3kb, 2.5kb and 1.5kb and very light smear above them. Tomorrow I need to send for sequencing this plasmid with 2 primers (that I already have) to verify once and for good the sequence! I hope for the BEST!!!<br> |
| | - Transformation of BBa_K346000 (T3 RNAP) from yesterday to the TOP10 bacteria. I plated 100ul and 200ul on LB+Amp plates. The plates are in 37C incubator for the night. | | - Transformation of BBa_K346000 (T3 RNAP) from yesterday to the TOP10 bacteria. I plated 100ul and 200ul on LB+Amp plates. The plates are in 37C incubator for the night. |
| | ==Asaf== | | ==Asaf== |
| - | | + | I ran a PCR reaction of the different plasmids that contains the polymerase genes (T7*,N4,T3,K1F,SP6)<br> |
| | + | in order to extract these genes. |
| | ==Hila== | | ==Hila== |
| | + | Gel purification for fragments 2, 3, 4, 5, 7, 8 (with and without restriction sites). <br> |
| | | | |
| | ==Lior== | | ==Lior== |
| | | | |
| | ==Noa== | | ==Noa== |
| | + | Transformation of ALP + N4 T3 K1F Sp6, Xy + N4 T3 K1F Sp6. <br> |
| | + | Plating on CM plates. |
| | | | |
| | ==Evgeni== | | ==Evgeni== |
| | + | - Made miniprep and restriction analysis from the new starters again. Still no positives. I will try to do ligation again, however this time I'll treat a pPROLar backbone with CIP to prevent self ligation, will raise insert:plasmid ratio to 5:1 instead of 3:1 in previous ligation and put ligation overnight instead one hour in 30 degrees. |
| | + | <br> |
| | | | |
| | + | - Digestion of pPROLar again with BamHI-HF and HindIII-HF + cleaning for the new ligation. |
| | + | |
| | + | |
| | ==Shahar== | | ==Shahar== |
| - | | + | Miniprep to pSB1C3+MCS(1) & pSB1C3+MCS(3) |
| | ==Rachel== | | ==Rachel== |
| | | | |
| | }} | | }} |
Latest revision as of 14:38, 5 September 2012
Team:Technion/28_August_2012
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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/wiki/index.php?title=Team:Technion/4_May_2012&preload=Team:Technion/Template:NewDate&action=edit 4]
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| [http://2012.igem.org/Team:Technion/10_May_2012 10]
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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/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/26_July_2012 26]
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| [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]
|
| [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]
|
| [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
- Put starters of pSB1C3+Fus2, pSB1C3+Fus2del, pSB3C5+Fus2, pSB3C5+Fus2del, pSB1C3+MCS for the plate reader tomorrow.
Inbal
-I did a research on the SP6 gene that was sent to us from china- I found the sequence of the plasmid it came with, I also found the map+restriction sites of the plasmid (pACSP6R, or his other commercial name: pACYC184).
after that, I digested this plasmid with BamHI+HindIII and ran on 1% agarose gel. The expected bands were: 5600 and 350 bp approximately. The actual bands that I got were 3 light bands: 3kb, 2.5kb and 1.5kb and very light smear above them. Tomorrow I need to send for sequencing this plasmid with 2 primers (that I already have) to verify once and for good the sequence! I hope for the BEST!!!
- Transformation of BBa_K346000 (T3 RNAP) from yesterday to the TOP10 bacteria. I plated 100ul and 200ul on LB+Amp plates. The plates are in 37C incubator for the night.
Asaf
I ran a PCR reaction of the different plasmids that contains the polymerase genes (T7*,N4,T3,K1F,SP6)
in order to extract these genes.
Hila
Gel purification for fragments 2, 3, 4, 5, 7, 8 (with and without restriction sites).
Lior
Noa
Transformation of ALP + N4 T3 K1F Sp6, Xy + N4 T3 K1F Sp6.
Plating on CM plates.
Evgeni
- Made miniprep and restriction analysis from the new starters again. Still no positives. I will try to do ligation again, however this time I'll treat a pPROLar backbone with CIP to prevent self ligation, will raise insert:plasmid ratio to 5:1 instead of 3:1 in previous ligation and put ligation overnight instead one hour in 30 degrees.
- Digestion of pPROLar again with BamHI-HF and HindIII-HF + cleaning for the new ligation.
Shahar
Miniprep to pSB1C3+MCS(1) & pSB1C3+MCS(3)
Rachel
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