09/09/13
From 2013.igem.org
(Difference between revisions)
Line 33: | Line 33: | ||
|- | |- | ||
|TOD X6||31.7||70.5||1.88||1.88 | |TOD X6||31.7||70.5||1.88||1.88 | ||
+ | |- | ||
+ | | | ||
+ | |TOD F1||32.4||39.6||1.91||1.59 | ||
+ | |- | ||
+ | |TOD F2||33.7||49.4||1.91||1.65 | ||
+ | |- | ||
+ | |TOD F3||32.3||47.4||1.89||1.71 | ||
+ | |- | ||
+ | |TOD F4||31.6||43.7||1.94||1.73 | ||
+ | |- | ||
+ | |TOD F5||36.5||57||1.89||1.50 | ||
+ | |- | ||
+ | |TOD F6||38||59.4||1.89||1.61 | ||
+ | |} | ||
+ | |||
|} | |} | ||
Revision as of 16:35, 9 September 2013
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Miniprep of overnight broth
- Following on from 06/09/13 result,overnight broth was prepared yesterday from the single colonies that had grown on each of the 2 plates shown on 06/09/13.
- The broth, which contains cells that have been transformed with pSB1C3/TOD genes, was minipreped using the Omega Bio-Tek kit.
- This would enable us to confirm which cells have the recircularised plasmid and those with the ligated product.
Measurement of DNA concentration
Samples | Volume (ul) | Conc (ng/ul) | 280/260 | 260/280 | |
TOD X1 | 31.6 | 86.5 | 1.87 | 1.92 | |
TOD X2 | 31.7 | 58.7 | 1.89 | 1.26 | |
TOD X3 | 32.7 | 56.6 | 1.91 | 1.88 | |
TOD X4 | 31.6 | 55.8 | 1.88 | 1.65 | |
TOD X5 | 31.7 | 44.4 | 1.87 | 1.78 | |
TOD X6 | 31.7 | 70.5 | 1.88 | 1.88 | |
TOD F1 | 32.4 | 39.6 | 1.91 | 1.59 | |
TOD F2 | 33.7 | 49.4 | 1.91 | 1.65 | |
TOD F3 | 32.3 | 47.4 | 1.89 | 1.71 | |
TOD F4 | 31.6 | 43.7 | 1.94 | 1.73 | |
TOD F5 | 36.5 | 57 | 1.89 | 1.50 | |
TOD F6 | 38 | 59.4 | 1.89 | 1.61 |
|}
Double Digestion of pSB1C3/TOD genes
- The pSB1C3/TOD genes were double digested with (XbaI and SpeI) & (EcoRI and PstI)
- The master mix for the (XbaI and SpeI) was;
60ul of cut smart buffer 5ul of XbaI 10ul of SpeI 273ul of 5M Tris
- The master mix for the (EcoRI and PstI) was;
60ul of cut smart buffer 5ul of XbaI 5ul of SpeI 278ul of 5M Tris
All 36 tubes were incubated overnight in 37 degrees room.
- Tomorrow the plasmids that were isolated from the miniprep would be run on a 1% agarose gel to distinguish between cells that have the pSB1C3/TOD genes from those that have the recircularised pSB1C3 vector.