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09/09/13
From 2013.igem.org
(Difference between revisions)
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==Measurement of DNA concentration== | ==Measurement of DNA concentration== | ||
| - | == | + | |
| - | * The plasmids that were isolated from the miniprep | + | ==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. | ||
Revision as of 16:13, 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
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.
