From 2012.igem.org
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| | ==Inbal== | | ==Inbal== |
| | + | -I ran the rest of T7* RNAP with pSB1A2 primers on agarose gel, but the band was too faded to purify it (still- I'm going to purify it, just in case...) so I repeat the PCR twice for it but now I changed the Tm, one reaction was Tm=54C, the other was Tm=56C (After consultation with Shachar and roee, we decided to change the Tm's to gradient tempratures). After running a 5ul sample from the PCR products, I got the desired bands (2700bp), with no other unwanted bands, So I purified the products by PCR cleaning kit. The concentrations are: for 54C:207ng/ul, for 56C: 298ng/ul.<br> |
| | + | - 3 starters of k133 (the inserts are pTetO+m-cherry) that I received from Roee's lab. |
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| | ==Asaf== | | ==Asaf== |
| - | | + | I ran the PCR products of the polymerases on an extracting gel. I got the expected results (although N4 is rather week).<br> |
| | + | <gallery perrow="4" widths=150px heights=150px style="color: black; border: 5px ridge #A7A3BF; font: 13px david; background: azure"> |
| | + | Image:2.9_polymerase_extraction_gel-view.jpg |
| | + | </gallery> |
| | + | |
| | ==Hila== | | ==Hila== |
| | I wanted to restrict the fragments with PacI and SpeI, but… <br> | | I wanted to restrict the fragments with PacI and SpeI, but… <br> |
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| | ==Shahar== | | ==Shahar== |
| - | | + | PCR for N4 at 62 & 60 degrees, and for T3 worked with pPROLAR primers , but not with pTETO primers. <br> Digest of the RNAP with HindIII and NHeI for creating sticky ends |
| | ==Rachel== | | ==Rachel== |
| - | | + | I ran the PCR products of K1F RNAP (which designated for cloning into pPROLAR)on an extracting gel. <br> The result was as i expected for (2700 bp). |
| | }} | | }} |
Latest revision as of 08:48, 20 September 2012
Team:Technion/2_September_2012
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| [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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| 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
Inbal
-I ran the rest of T7* RNAP with pSB1A2 primers on agarose gel, but the band was too faded to purify it (still- I'm going to purify it, just in case...) so I repeat the PCR twice for it but now I changed the Tm, one reaction was Tm=54C, the other was Tm=56C (After consultation with Shachar and roee, we decided to change the Tm's to gradient tempratures). After running a 5ul sample from the PCR products, I got the desired bands (2700bp), with no other unwanted bands, So I purified the products by PCR cleaning kit. The concentrations are: for 54C:207ng/ul, for 56C: 298ng/ul.
- 3 starters of k133 (the inserts are pTetO+m-cherry) that I received from Roee's lab.
Asaf
I ran the PCR products of the polymerases on an extracting gel. I got the expected results (although N4 is rather week).
Hila
I wanted to restrict the fragments with PacI and SpeI, but…
NO PacI!!! :(
Lior
Noa
Colony PCR for: ALP + k1F, Sp6.
Miniprep and Nanodrop for all: xyIE + all, and ALP + N4, T3.
Evgeni
- For some reason I got no growth in starters, though there was growth on streak plate. Will try to make starters again, this time from the colonies on streak plate.
Shahar
PCR for N4 at 62 & 60 degrees, and for T3 worked with pPROLAR primers , but not with pTETO primers.
Digest of the RNAP with HindIII and NHeI for creating sticky ends
Rachel
I ran the PCR products of K1F RNAP (which designated for cloning into pPROLAR)on an extracting gel.
The result was as i expected for (2700 bp).
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