Difference between revisions of "Team:METU Turkey/Experiments"
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<h2 align=center>Experiments/Documentation of the development of our project</h2> | <h2 align=center>Experiments/Documentation of the development of our project</h2> | ||
− | <p align=center> | + | <p align=center>Standard iGEM <a class="one" href="http://parts.igem.org/Help" target="_blank">Protocols</a></p> |
− | </p> | + | <pre> |
− | </div> | + | |
+ | July 15th, Wednesday | ||
+ | Stock cells were planted in empty plates | ||
+ | |||
+ | July 16th, Thursday | ||
+ | Competent cells were prepared | ||
+ | 7 strains | ||
+ | Transformation was done at night for control purposes, 5 strains were negative (amp plate 4 min 4000 rpm) | ||
+ | |||
+ | July 17th, Friday | ||
+ | Kitten gene was extracted | ||
+ | 5 strains | ||
+ | Transformation was done for control purposes. This was so because 5 strains were negative in the previous one. | ||
+ | Kumomax enzyme was planted from agar plate to agar plate. | ||
+ | |||
+ | July 18th, Saturday | ||
+ | All of the transformations were positive. | ||
+ | Kumomax enzyme was planted from agar to LB. | ||
+ | |||
+ | July 19th, Sunday | ||
+ | Plasmid isolation was done. | ||
+ | Digestion was done (with not1) | ||
+ | Gel electrophoresis was done, The result was positive. | ||
+ | Gel extraction was done | ||
+ | Nanodrop result was 77ng/microl | ||
+ | |||
+ | July 20th, Monday | ||
+ | Plate with zeocin prepared (salt 5g,) total 200 mL | ||
+ | Low salt medium (LB) was prepared, total 200 mL | ||
+ | |||
+ | July 21st, Tuesday | ||
+ | Transformation was done. PGapZ -alpha was duplicated. | ||
+ | |||
+ | July 22nd, Wednesday | ||
+ | PGapZ-alpha was planted from agar to LB | ||
+ | |||
+ | July 23rd, Thursday | ||
+ | Plasmid isolation was done for PGapZ-alpha | ||
+ | Restriction was done with NotI (PGapZ-alpha) | ||
+ | Gel electrophoresis was done (PgapZ-alpha), the result was positive | ||
+ | Geş extraction was done, the nanadrop result was positive | ||
+ | |||
+ | July 24th, Friday | ||
+ | Ligation was done for PGapZ-alpha-kumomax | ||
+ | Transformation was done | ||
+ | |||
+ | July 25th, Saturday | ||
+ | Transformation results were negative | ||
+ | Transformation was done | ||
+ | 323101 chl | ||
+ | 323106 chl | ||
+ | 323117 chl | ||
+ | I13504 (GFP) Amp | ||
+ | 8 transformation plates total, two plates from each. | ||
+ | |||
+ | July 26th, Sunday | ||
+ | Transformation results were positive | ||
+ | The colonies were taken from agar and planted in LB | ||
+ | |||
+ | July 27th, Monday | ||
+ | Plasmid isolation was done for the promoter in Lb and GFP | ||
+ | Restriction was done | ||
+ | For promoters, (101, 106, 117) Spe1 and pst1 | ||
+ | For GFP, Xba1 and PSt 1 | ||
+ | Gel electrophoresis ws done for the cut promoters and GFP | ||
+ | 101, 106, 117 and Gfp were again planted to LB from agar | ||
+ | |||
+ | July 28th, Tuesday | ||
+ | Plasmid isolation was done for 101, 106, 117 and GFP | ||
+ | Restriction was done | ||
+ | Gel electrophoresis was done for the cut 101, 106, 117 and GFP, the results were negative | ||
+ | 101, 106, 117 and GFP were again planted in LB from agar | ||
+ | |||
+ | July 29th, Wednesday | ||
+ | Plasmid isolation was done for 101, 106, 117 and GFP | ||
+ | Restriction was done for 101, 106, 117 and GFP | ||
+ | Gel electrophoresis was done for the cut 101, 106, 117 and GFP, the results were positive only for the promoters, 101, 106, and 117 | ||
+ | Gel extraction was done for promoters | ||
+ | Nanodrop results were low | ||
+ | 101, 106, 117 and GFP were again planted in LB from agar | ||
+ | |||
+ | July 30th, Thursday | ||
+ | Plasmid isolation was done for 101, 106, 117 and GFP | ||
+ | Centrifuge was lowered to 12000 rpm from 13000 rpm | ||
+ | Restriction was done for 101, 106, 117 and GFP | ||
+ | Optimization was done for restriction | ||
+ | |||
+ | |||
+ | <img hspace="375px" align=center src="https://static.igem.org/mediawiki/2015/2/2b/4_sayfa_elif_tablo.PNG" width="600px"> | ||
+ | |||
+ | |||
+ | |||
+ | The electrophoresis results were acquired for set 2 and set 2, set 1 did not produce results | ||
+ | Gel extraction was done for the 4 bands that yielded positive results | ||
+ | Nanodrop results were low | ||
+ | Plantation was done again from agar to LB | ||
+ | |||
+ | July 31st, Friday | ||
+ | Plasmid isolation as done for GFP | ||
+ | Restriction was done by Xba1 and Pst1 for GFP | ||
+ | Gel electrophoresis was done, the results were positive | ||
+ | Gel extraction was done | ||
+ | Nanodrop results were low | ||
+ | Plantation from agar to LB was done again for GFP | ||
+ | |||
+ | August 1st, Saturday | ||
+ | Plasmid isolation was done for GFP | ||
+ | |||
+ | August 3rd, Monday | ||
+ | Restriction was done for GFP | ||
+ | Gel electrophoresis was performed on the ones that were cut | ||
+ | The results were positive | ||
+ | Gel extraction was performed | ||
+ | Nanodrop results were good | ||
+ | |||
+ | August 4th, Tuesday | ||
+ | Ligation was done for promoters and GFP | ||
+ | |||
+ | August 5th, Wednesday | ||
+ | Transformation results were negative | ||
+ | Ligation was performed again | ||
+ | Transformation was done for those who had been ligated | ||
+ | |||
+ | August 6th, Thursday | ||
+ | Transformation results are negative | ||
+ | Ligation was performed again for promoters and GFP | ||
+ | Transformation was done | ||
+ | Ligation was done for PGapZ-alpha and kumomax | ||
+ | Transformation was done | ||
+ | |||
+ | August 7th, Friday | ||
+ | The transformations done for promoter-GFP and PGapZ-alpha-kumomaz were negative | ||
+ | 400 mL Amp agar medium was prepared, put into plates | ||
+ | |||
+ | August 10th, Monday | ||
+ | Streak plate was done for 101, 106, 117 and GFP (8 in total) | ||
+ | Chl plates were prepared | ||
+ | -400 mL agar medium | ||
+ | -5 g salt was used instead of 10 g salt | ||
+ | |||
+ | August 11th, Tuesday | ||
+ | The streaked plated 101, 106,117 and GFP were planted in LB (8 tubes in total) | ||
+ | LB was prepared (400 mL, 5 g salt) | ||
+ | |||
+ | August 12th, Wednesday | ||
+ | Plasmid isolation was done for 101, 106, 117 and GFP | ||
+ | Restriction was done for GFP | ||
+ | <img hspace="375px" align=center src="https://static.igem.org/mediawiki/2015/3/3a/7_son_sayfa_elif_tablo.PNG" width="600px"> | ||
+ | |||
+ | </pre> | ||
+ | </div></br> | ||
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Revision as of 19:57, 17 September 2015
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