From 2012.igem.org
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| | - Glycerol stock of pSB1C3+Fus2del (freezer position G2) and pSB3C5+Fus2del (freezer position G3).<br> | | - Glycerol stock of pSB1C3+Fus2del (freezer position G2) and pSB3C5+Fus2del (freezer position G3).<br> |
| | - Plate reader assay for pSB1C3+Fus2 and pSB3C5+Fus2 with the positive (pSB1C3+Fus2del and pSB3C5+Fus2del) and negative (pSB1C3+MCS) controls. | | - Plate reader assay for pSB1C3+Fus2 and pSB3C5+Fus2 with the positive (pSB1C3+Fus2del and pSB3C5+Fus2del) and negative (pSB1C3+MCS) controls. |
| - | | + | |
| | ==Inbal== | | ==Inbal== |
| | + | Transformation for pACSP6R and for BBa_K346000 (native T3 RNAP) to TOP10 bacteria, the resistances were Tetracycline and kanamycin respectively. Then, I plated on plates and incubate them in 37C incubator overnight.<br> |
| | + | Also, I did 2 PCR reactions for T7* RNAP that we got from Chris, each reaction with different primers ( one primer compatible for the pSB1A2 plasmid, the second is compatible for pPROLAR plasmid). The Tm was 58C, according to Roee's guidance. |
| | + | After the PCR, I ran 5ul in 1% agarose gel, and I saw a desired 2700bp sized band with the pPROLAR primers, but no bands in the pSB1A2 primers. I don't know what else to do :/ |
| | + | I digested the pACSP6R plasmid with BamHI-HF enzyme- the results: good :] - after running it on gel, I expected a 5.9kb band, and that's what I saw in the gel! finally an appropriate result!!! |
| | | | |
| | ==Asaf== | | ==Asaf== |
| - | | + | -I checked the results of yesterday's PCR for extracting the polymerase genes on gel agarose,<br> |
| | + | and got two bands in T7*,T3 and K1f: one in 2kb and one near 3kb, and no bands in SP6 and N4. <br> |
| | + | I realized that I put an incorrect elongation time and this can explain the double band.<br> |
| | + | The Sp6 gene was probably defected.<br> |
| | + | -I ran another PCR reaction for the polymerase genes. This time with the right elongation time, but with Tm of 60C.<br> |
| | + | I didn't got any bands with the exception of T3. |
| | + | |
| | ==Hila== | | ==Hila== |
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| | ==Noa== | | ==Noa== |
| | + | Colony PCR for ALP bacteria described yesterday. <br> |
| | + | Positive results for: ALP+ T3 , N4. |
| | | | |
| | ==Evgeni== | | ==Evgeni== |
| | + | - Overnight ligation of pPROLar backbone with Bba_015 |
| | | | |
| - | ==Shahar== | + | ==Rachel & Shahar== |
| | PCR to Chris's Rnap with the new primers. The PCR with plac/ARA primer and T3, K1F & T7* templates, were successful. | | PCR to Chris's Rnap with the new primers. The PCR with plac/ARA primer and T3, K1F & T7* templates, were successful. |
| | The PCR with A@ primer and T3, K1F, N4 & T7* templates, failed. <br> | | The PCR with A@ primer and T3, K1F, N4 & T7* templates, failed. <br> |
| - | ==Rachel==
| + | |
| | | | |
| | }} | | }} |
Latest revision as of 12:20, 6 September 2012
Team:Technion/29_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/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/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]
| [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/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/16_June_2012&preload=Team:Technion/Template:NewDate&action=edit 16]
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| [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/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]
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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/Team:Technion/4_August_2012 4]
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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]
|
| [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
- Glycerol stock of pSB1C3+Fus2del (freezer position G2) and pSB3C5+Fus2del (freezer position G3).
- Plate reader assay for pSB1C3+Fus2 and pSB3C5+Fus2 with the positive (pSB1C3+Fus2del and pSB3C5+Fus2del) and negative (pSB1C3+MCS) controls.
Inbal
Transformation for pACSP6R and for BBa_K346000 (native T3 RNAP) to TOP10 bacteria, the resistances were Tetracycline and kanamycin respectively. Then, I plated on plates and incubate them in 37C incubator overnight.
Also, I did 2 PCR reactions for T7* RNAP that we got from Chris, each reaction with different primers ( one primer compatible for the pSB1A2 plasmid, the second is compatible for pPROLAR plasmid). The Tm was 58C, according to Roee's guidance.
After the PCR, I ran 5ul in 1% agarose gel, and I saw a desired 2700bp sized band with the pPROLAR primers, but no bands in the pSB1A2 primers. I don't know what else to do :/
I digested the pACSP6R plasmid with BamHI-HF enzyme- the results: good :] - after running it on gel, I expected a 5.9kb band, and that's what I saw in the gel! finally an appropriate result!!!
Asaf
-I checked the results of yesterday's PCR for extracting the polymerase genes on gel agarose,
and got two bands in T7*,T3 and K1f: one in 2kb and one near 3kb, and no bands in SP6 and N4.
I realized that I put an incorrect elongation time and this can explain the double band.
The Sp6 gene was probably defected.
-I ran another PCR reaction for the polymerase genes. This time with the right elongation time, but with Tm of 60C.
I didn't got any bands with the exception of T3.
Hila
Lior
Noa
Colony PCR for ALP bacteria described yesterday.
Positive results for: ALP+ T3 , N4.
Evgeni
- Overnight ligation of pPROLar backbone with Bba_015
Rachel & Shahar
PCR to Chris's Rnap with the new primers. The PCR with plac/ARA primer and T3, K1F & T7* templates, were successful.
The PCR with A@ primer and T3, K1F, N4 & T7* templates, failed.
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