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Team:Fatih Turkey/Notebook8
From 2011.igem.org
04.09.11
1. For each815, 815 gex, 825, 825 gex, 826, 926, 915, 915 gex, 925, 925 gex two colonies had been chosen and all were found wrong; thus this procedure is replied.
2. The parts above are isolated from their LBs.
3. Same parts are then digested with Xba1 and Pst1.
4. Same parts are loaded into gel.
a. For 915 and 915 gex, destinationplasmids were foundwrong; but genes were correct.
b. We were not so sure for destination plasmid of 826; but its gene were also correct.
https://static.igem.org/mediawiki/2011/2/2a/04.09.11.png
https://static.igem.org/mediawiki/2011/4/45/04%2C09%2C2011.pdf
05.09.11
https://static.igem.org/mediawiki/2011/7/7a/05.09.11_-_II.png
https://static.igem.org/mediawiki/2011/5/56/05%2C09%2C11_DO%C4%9ERUSU.pdf
https://static.igem.org/mediawiki/2011/7/7a/05.09.11_-_II.png
06.09.11
1. 815 ep, 815 xp, 915 ep, 915 xp, 915 gexxp, 515 xparedigested-ligated. (Note that transformation is not performed because of absence of prepared compotent cell.)
2. 545, 504-GFP and 915 aretransformated.
3. Each for 596, 596-b, 546, 536; two colonies are chosen. LBs for these parts are prepared. (8 LBs)
4. For 815, 815 gex, 915 and 915 gex; four additional LBs are prepared in order to gain more copies of them.
5. An antibiotic free LB is prepared to make compotent cell next day. One compotent cell in the stock is transferred into it.
https://static.igem.org/mediawiki/2011/b/b1/06%2C09%2C2011.pdf
https://static.igem.org/mediawiki/2011/4/4d/916_e%2Cp.pdf
07.09.11
1. We made plasmid isolation to 536H,546H,596B_G,596B_H,596H , j04500 but unfortunatelly we fixed 596 G and 546 G in experiment time .With xba1 and pst1:
536H >>>> 13.1 29.4
546H >>>>> 7.4 35.1
596B_G >>>> 15.1 27.4
596B_H >>>> 12.5 30
596G+546>>>> 11.6 30.9
596 H >>>> 16.1 26.4
With ecor1 and spe1:
J04500_2(double dig.for up) >>>> 6.6 35.9
With e.cor1:
J04500_2(single)>>>> 6.6 36.9
J04500_2(undigest)>>>> 6.6 37.9(WE FORGOT THİS!!! So we didnt made electrophoresis j04500 _2 single and undigest and didnt put all of j04500 _2(double dig.for up ) to gel.
2.then made them electrophoresis.
3. 815 ,815 gex,545,916,j04450,can are put liquid culture from glyserol stock. We choose 2 colonies from 915 gx xp and also 2 colonies from 504 gfp then put all of them 10 ml liquid culture.
4.Top 10,DH 5alfa ,PET 28 a in BL21,BL 21 are put 10 ml liquid culture.
https://static.igem.org/mediawiki/2011/1/1b/07%2C09%2C2011.confirmed.pdf
https://static.igem.org/mediawiki/2011/e/e5/07%2C09%2C2011.pdf
https://static.igem.org/mediawiki/2011/f/fe/07.09.11.png
08.09.11
1. For each of 915 gexxpand 504-gfp, two colonies were incubated in 10 ml LB.
2. 916, 900, 915 gexxp, 504-gfp, 545, 500, 815, 815 gex are isolated.
3. The parts above are digested with Xba1 and Pst1 except 545, 500.The separts are remainedun digested; because both of them had been confirmed.
4. Digested parts are loaded into gel.
a. All parts except 900 are found wrong.
https://static.igem.org/mediawiki/2011/2/2b/08.09.11.png
https://static.igem.org/mediawiki/2011/e/e2/08%2C09%2C2011.pdf
09.09.11
1. Toligate 816, 825, 826, 916, 925, 926, 536 and 546; upstream, downstream and destination plasmids are digested with appropiate enzymes.
2. Digested solutions are loaded into gel and all of them are confirmed.
3. For each of 815 and 915 parts in the stock; LB solutions are prepared.
https://static.igem.org/mediawiki/2011/3/36/09.09.11.png
10.09.11
1. Because of absence of 800, digested solutions that prepared for the ligation of 816, 825 and 826 are notligated.
2. 536, 546, 916, 925 and 926 are ligated.
3. 916, 925 and 926 transformated twice. (We have two plates each of these parts.) 536 and 546 are transformated into only one plate.
4. LBs that were prepared yesterday are prepared again. (Isolation could not take place today.)
11.09.11
https://static.igem.org/mediawiki/2011/a/a5/11%2C09%2C2011_2.pdfhttps://static.igem.org/mediawiki/2011/0/0d/11%2C09%2C2011.pdf
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