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Template:Team:KULeuven/1 September 2009/VanillinProduction
From 2009.igem.org
(Difference between revisions)
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* SAMS I B RD is used for ligation with TER II RD | * 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 | + | ** 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 | + | ** The restriction enzymes are denatured by keeping the sample at 85°C for 20 minutes |
{| border ="1" align="center" | {| border ="1" align="center" | ||
!| vector || insert | !| vector || insert | ||
| Line 33: | Line 33: | ||
|- | |- | ||
|} | |} | ||
| - | * Results on gel | + | * 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. | * 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... | ** No signal... grrr... | ||
* 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, | + | # 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. |
Revision as of 06:35, 3 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 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.
