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09/09/13
From 2013.igem.org
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Contents |
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 |
| TOB G1 | 36.8 | 72.5 | 1.89 | 1.51 |
| TOB G2 | 44.7 | 1.90 | 1.90 | 1.65 |
| TOB G3 | 33.7 | 50.2 | 1.90 | 1.46 |
| TOB G4 | 41.9 | 55 | 1.93 | 1.35 |
| TOB G5 | 34 | 43 | 1.96 | 1.54 |
| TOB G6 | 35 | 48 | 1.89 | 1.72 |
DNA concentration was measured using Nanodrop.
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.
Dissolution of polystyrene in Limonene
- This experiment was set up to decide how much limonene can economically dissolve polystyrene.
MATERIALS USED 20ml solution of 25% limonene. 8cm x 5cm x 2cm polystyrene block (thus volume of 80cm^3)
- It took ~33 minutes for the block to be completely dissolved
