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Template:Team:KULeuven/1 September 2009/VanillinProduction
From 2009.igem.org
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* Replated colonies from the original SAMS plate from 24/8 | * Replated colonies from the original SAMS plate from 24/8 | ||
** Took 4 individual colonies and plated them on 4 corners of a petri dish | ** Took 4 individual colonies and plated them on 4 corners of a petri dish | ||
| + | # the 4 genes(sam5, sam8, ech, fcs) where taken from the original DNA that was sent, was electroporated and plated. this in order to make a glycerol stock. | ||
Revision as of 13:00, 2 September 2009
- SAMS I B RD is used for ligation with TER II RD
- Since we want to avoid gel purification we use the direct sample from the restriction diget
- The restriction enzymes are denatured by keeping the sample at 85C for 20 minutes
| vector | insert |
|---|---|
| Terminator | SAMS |
| 2000 bp | 3070 bp |
| 50 ng | 280 ng |
| 10 μl | 16 μl |
- The genes fcs and ech are individually cut with the four restriction enzymes
| EcoR1 | fcs RD-E |
| Spe1 | fcs RD-S |
| Pst1 | fcs RD-P |
| Xba1 | fcs RD-X |
| EcoR1 | Ech RD-E |
| Spe1 | Ech RD-S |
| Pst1 | Ech RD-P |
| Xba1 | Ech RD-X |
- Results on gel shows that all the genes have the same length and show 1 line.
- SAMS I A RD is put on gel, to be used for gel extraction and a part of SAMS I B RD is put on gel to check the length.
- No signal... grrr...
- Replated colonies from the original SAMS plate from 24/8
- Took 4 individual colonies and plated them on 4 corners of a petri dish
- the 4 genes(sam5, sam8, ech, fcs) where taken from the original DNA that was sent, was electroporated and plated. this in order to make a glycerol stock.
