Template:Team:KULeuven/1 September 2009/VanillinProduction
From 2009.igem.org
- 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 digest
- The restriction enzymes are denatured by keeping the sample at 85°C 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 show that all the genes have the same length and display 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, electroporated and plated. This in order to make a glycerol stock.