Team:Technion/11 September 2012

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- Did a restriciton digest with BamHI-HF to test the colonies for the correct insert. 3/3 clones of Fus2del and 3/4 clones of Fus2 were positive. I picked one of each of the clones for plate reader tomorrow.<br>
- Did a restriciton digest with BamHI-HF to test the colonies for the correct insert. 3/3 clones of Fus2del and 3/4 clones of Fus2 were positive. I picked one of each of the clones for plate reader tomorrow.<br>
- Put starters for plate reader tomorrow.<br>
- Put starters for plate reader tomorrow.<br>
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- Tested ladders on a 0.6% agarose gel. Ran for an hour and 10 mins at 40V on a minigel. The 1Kb opened but the high range (HR) ladder didn't open. We will a 0.4% agarose gel in the future attempts.<br>
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- Tested ladders on a 0.6% agarose gel. Ran for an hour and 10 mins at 40V on a minigel. The 1Kb opened but the high range (HR) ladder didn't open. We will use a 0.4% agarose gel in the future attempts.<br>
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- Add gibson description and pic.
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- Hila and I did the first attempt for gibson on the phage fragments. We attempted an assembly of fragments #4 (8080bp) and #5 (6150bp). The results are presented in the following gel pic: <br>
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[[File:Two parts together 11.9.12.jpg|350px]]<br>
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As you can see in the attached file, only one of our three attempts today actually worked. The reaction conditions for the attempt that worked were: 100bp overlaps, ~150ng DNA of each of the templates (~0.025-0.03 pmoles), 5ul of Gibson mix and 5 ul of DNA fragments (10ul final volume). All reactions were incubated for 1 hr at 50 degrees as instructed by the manual.<br>
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I think that the 20 ul final volume reaction with the 100bp didn't work because there was too much DNA. The 400bp overlap might still work with additional exonuclease and in a 10ul volume with ~150ng of each fragment.
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==Inbal==
==Inbal==
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-Miniprep for two samples out of three from the starters I made yesterday- because one of them has been contaminated. The conc's are: 146.5 and 142 ng/ul. I didn't know for sure if the insert (eT7) is within the plasmid (pSB1AK3) so I digested the plasmid with EcoRI. After running the products on a gel, comparison to the uncut plasmid, I got the desired band- 6kb approximately! The clone works! <br>
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- Shachar helped me to do a second transformation of native T3 to pSB1AK3 (2 transformations of 10ul and 6 ul from the ligation), we plated on 4 amp resistance plates (100ul and rest) and incubate them overnight at 37C. <br>
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- restriction of eT7+pSB1AK3 with XbaI, then I cleaned the products from buffers,etc.. and continue it to a cip reaction,  after that I measured the concentration: 13.2 ng/ul. I ligated an inducible promoter (pTetO) attached to a reporter gene (m-cherry) to eT7+pSB1AK3, 37C for 1 hour.<br>
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- I transformed the ligation product into TOP10 component cells +control of cip.<br>
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- Starter for eT7+pSB1AK3 for cloning it to shipping plasmid.<br>
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==Asaf==
==Asaf==
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I cleaned the DNA from the extracted gel. I got a very low DNA concentration.
==Hila==
==Hila==
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- Gel purification for F4 (400bp overlap), F4*(100bp overlap) and F5. <br>
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- Colony PCR for additional five colonies of fragments 4C, 5C and 7C. I found one positive colony for fragment 5. <br>
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- Transformation of ligated 1C and 6C to Top10-Rb bacteria. <br>
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- Starter for pBS1C3_MCS and for the positive colonies with the plasmid containing fragments 2C, 3C. 5C and 8C. <br>
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- Additional amplification for F4, F4* and F5. <br>
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- First Gibson assembly attempt. For more information – please read what Ilya wrote… <br>
==Lior==
==Lior==
-
 
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The trasformation worked, we have T7 promoter parts in a bacteria with PET system. <br>
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In order to test the system we used various concentrations of IPTG and checked absorbance for the ALP and xyIE part. <br>
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We also did coloby PCR for the positive control parts and made starters for the positive ones.
==Noa==
==Noa==
==Evgeni==
==Evgeni==
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Transformation of ligation products into TOP10 ''E. coli'' strain
==Shahar==
==Shahar==
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after using the PCR products of N4 & T3 with pPROLAR primers as template - we needed to do another PCR to produce new N4 & T3 templates with pPROLAR prefix and sufix.<br>
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helped with colony PCR for additional five colonies of fragments 4C, 5C and 7C. <br>
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helped with transformation of ligated 1C and 6C to Top10-Rb bacteria. 
==Rachel==
==Rachel==
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PCR for pTetO+m-cherry , after that  i ran on gel the PCR products and got the band i expected for:800bp, then I cleaned the PCR products. The concentration was: 119 ng/ul. <br>
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Digestion of pTetO+m-cherry with xbaI & speI to create sticky ends. cleaning after restriction, the concentration was 63 ng/ul.
}}
}}

Latest revision as of 18:42, 26 September 2012



Team:Technion/11_September_2012
March
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[http://2012.igem.org/wiki/index.php?title=Team:Technion/5_March_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_March_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_March_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_March_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_March_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_March_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_March_2012&preload=Team:Technion/Template:NewDate&action=edit 11]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/12_March_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_March_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_March_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_March_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_March_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_March_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_March_2012&preload=Team:Technion/Template:NewDate&action=edit 18]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/19_March_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_March_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/Team:Technion/21_March_2012 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_March_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_March_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_March_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_March_2012&preload=Team:Technion/Template:NewDate&action=edit 25]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/26_March_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_March_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_March_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_March_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_March_2012&preload=Team:Technion/Template:NewDate&action=edit 30] [http://2012.igem.org/wiki/index.php?title=Team:Technion/31_March_2012&preload=Team:Technion/Template:NewDate&action=edit 31]
April
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            [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_April_2012&preload=Team:Technion/Template:NewDate&action=edit 1]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/2_April_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/Team:Technion/3_April_2012 3] [http://2012.igem.org/Team:Technion/4_April_2012 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_April_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_April_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_April_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_April_2012&preload=Team:Technion/Template:NewDate&action=edit 8]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/9_April_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_April_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_April_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_April_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_April_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/Team:Technion/14_April_2012 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_April_2012&preload=Team:Technion/Template:NewDate&action=edit 15]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/16_April_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_April_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_April_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_April_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_April_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_April_2012&preload=Team:Technion/Template:NewDate&action=edit 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_April_2012&preload=Team:Technion/Template:NewDate&action=edit 22]
[http://2012.igem.org/Team:Technion/23_April_2012 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_April_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/Team:Technion/25_April_2012 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_April_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_April_2012&preload=Team:Technion/Template:NewDate&action=edit 29]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/30_April_2012&preload=Team:Technion/Template:NewDate&action=edit 30]
May
MTWTFSS
  [http://2012.igem.org/Team:Technion/1_May_2012 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_May_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_May_2012&preload=Team:Technion/Template:NewDate&action=edit 6]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/7_May_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_May_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_May_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_May_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_May_2012&preload=Team:Technion/Template:NewDate&action=edit 13]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/14_May_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_May_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_May_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_May_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_May_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_May_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_May_2012&preload=Team:Technion/Template:NewDate&action=edit 20]
[http://2012.igem.org/Team:Technion/21_May_2012 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_May_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_May_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_May_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_May_2012&preload=Team:Technion/Template:NewDate&action=edit 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_May_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_May_2012&preload=Team:Technion/Template:NewDate&action=edit 27]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/28_May_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/31_May_2012&preload=Team:Technion/Template:NewDate&action=edit 31]
June
MTWTFSS
        [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_June_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_June_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/Team:Technion/3_June_2012 3]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/4_June_2012&preload=Team:Technion/Template:NewDate&action=edit 4] [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]
[http://2012.igem.org/Team:Technion/11_June_2012 11] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_June_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_June_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/Team:Technion/17_June_2012 17]
[http://2012.igem.org/Team:Technion/18_June_2012 18] [http://2012.igem.org/Team:Technion/19_June_2012 19] [http://2012.igem.org/Team:Technion/20_June_2012 20] [http://2012.igem.org/Team:Technion/21_June_2012 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_June_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_June_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/Team:Technion/24_June_2012 24]
[http://2012.igem.org/Team:Technion/25_June_2012 25] [http://2012.igem.org/Team:Technion/26_June_2012 26] [http://2012.igem.org/Team:Technion/27_June_2012 27] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_June_2012&preload=Team:Technion/Template:NewDate&action=edit 30]
July
MTWTFSS
            [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_July_2012&preload=Team:Technion/Template:NewDate&action=edit 1]
[http://2012.igem.org/Team:Technion/2_July_2012 2] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_July_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/Team:Technion/8_July_2012 8]
[http://2012.igem.org/Team:Technion/9_July_2012 9] [http://2012.igem.org/Team:Technion/10_July_2012 10] [http://2012.igem.org/Team:Technion/11_July_2012 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_July_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [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] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_July_2012&preload=Team:Technion/Template:NewDate&action=edit 15]
[http://2012.igem.org/wiki/index.php?title=Team:Technion/16_July_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/Team:Technion/17_July_2012 17] [http://2012.igem.org/Team:Technion/18_July_2012 18] [http://2012.igem.org/Team:Technion/19_July_2012 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_July_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_July_2012&preload=Team:Technion/Template:NewDate&action=edit 21] [http://2012.igem.org/Team:Technion/22_July_2012 22]
[http://2012.igem.org/Team:Technion/23_July_2012 23] [http://2012.igem.org/Team:Technion/24_July_2012 24] [http://2012.igem.org/Team:Technion/25_July_2012 25] [http://2012.igem.org/Team:Technion/26_July_2012 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_July_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_July_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/Team:Technion/29_July_2012 29]
[http://2012.igem.org/Team:Technion/30_July_2012 30] [http://2012.igem.org/Team:Technion/31_July_2012 31]
August
MTWTFSS
    [http://2012.igem.org/Team:Technion/1_August_2012 1] [http://2012.igem.org/Team:Technion/2_August_2012 2] [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]
[http://2012.igem.org/Team:Technion/6_August_2012 6] [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]
[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]
[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]
[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]
September
MTWTFSS
          [http://2012.igem.org/Team:Technion/1_September_2012 1] [http://2012.igem.org/Team:Technion/2_September_2012 2]
[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]
[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]
October
MTWTFSS
[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]
[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]
[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]
[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]
[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]
November
MTWTFSS
      [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]
[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]
[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]
[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]
[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]


Ilya

- Minipreped the picked colonies for testing the clones of Fus2 and Fus2del in pCP.
- Did a restriciton digest with BamHI-HF to test the colonies for the correct insert. 3/3 clones of Fus2del and 3/4 clones of Fus2 were positive. I picked one of each of the clones for plate reader tomorrow.
- Put starters for plate reader tomorrow.
- Tested ladders on a 0.6% agarose gel. Ran for an hour and 10 mins at 40V on a minigel. The 1Kb opened but the high range (HR) ladder didn't open. We will use a 0.4% agarose gel in the future attempts.
- Hila and I did the first attempt for gibson on the phage fragments. We attempted an assembly of fragments #4 (8080bp) and #5 (6150bp). The results are presented in the following gel pic:
Two parts together 11.9.12.jpg
As you can see in the attached file, only one of our three attempts today actually worked. The reaction conditions for the attempt that worked were: 100bp overlaps, ~150ng DNA of each of the templates (~0.025-0.03 pmoles), 5ul of Gibson mix and 5 ul of DNA fragments (10ul final volume). All reactions were incubated for 1 hr at 50 degrees as instructed by the manual.
I think that the 20 ul final volume reaction with the 100bp didn't work because there was too much DNA. The 400bp overlap might still work with additional exonuclease and in a 10ul volume with ~150ng of each fragment.

Inbal

-Miniprep for two samples out of three from the starters I made yesterday- because one of them has been contaminated. The conc's are: 146.5 and 142 ng/ul. I didn't know for sure if the insert (eT7) is within the plasmid (pSB1AK3) so I digested the plasmid with EcoRI. After running the products on a gel, comparison to the uncut plasmid, I got the desired band- 6kb approximately! The clone works!
- Shachar helped me to do a second transformation of native T3 to pSB1AK3 (2 transformations of 10ul and 6 ul from the ligation), we plated on 4 amp resistance plates (100ul and rest) and incubate them overnight at 37C.
- restriction of eT7+pSB1AK3 with XbaI, then I cleaned the products from buffers,etc.. and continue it to a cip reaction, after that I measured the concentration: 13.2 ng/ul. I ligated an inducible promoter (pTetO) attached to a reporter gene (m-cherry) to eT7+pSB1AK3, 37C for 1 hour.
- I transformed the ligation product into TOP10 component cells +control of cip.
- Starter for eT7+pSB1AK3 for cloning it to shipping plasmid.


Asaf

I cleaned the DNA from the extracted gel. I got a very low DNA concentration.

Hila

- Gel purification for F4 (400bp overlap), F4*(100bp overlap) and F5.
- Colony PCR for additional five colonies of fragments 4C, 5C and 7C. I found one positive colony for fragment 5.
- Transformation of ligated 1C and 6C to Top10-Rb bacteria.
- Starter for pBS1C3_MCS and for the positive colonies with the plasmid containing fragments 2C, 3C. 5C and 8C.
- Additional amplification for F4, F4* and F5.
- First Gibson assembly attempt. For more information – please read what Ilya wrote…

Lior

The trasformation worked, we have T7 promoter parts in a bacteria with PET system.
In order to test the system we used various concentrations of IPTG and checked absorbance for the ALP and xyIE part.
We also did coloby PCR for the positive control parts and made starters for the positive ones.

Noa

Evgeni

Transformation of ligation products into TOP10 E. coli strain

Shahar

after using the PCR products of N4 & T3 with pPROLAR primers as template - we needed to do another PCR to produce new N4 & T3 templates with pPROLAR prefix and sufix.
helped with colony PCR for additional five colonies of fragments 4C, 5C and 7C.
helped with transformation of ligated 1C and 6C to Top10-Rb bacteria.

Rachel

PCR for pTetO+m-cherry , after that i ran on gel the PCR products and got the band i expected for:800bp, then I cleaned the PCR products. The concentration was: 119 ng/ul.
Digestion of pTetO+m-cherry with xbaI & speI to create sticky ends. cleaning after restriction, the concentration was 63 ng/ul.