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Team:Groningen/Notebook/29 September 2009
From 2009.igem.org
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===Metal Accumulation=== | ===Metal Accumulation=== | ||
| - | *'''Redo As(III) uptake experiment for ''E. coli'' WT (pSB1A2) and ArsR-RFP''' | + | *'''Redo [https://2009.igem.org/Team:Groningen/Notebook/25_September_2009 As(III) uptake experiment] for ''E. coli'' WT (pSB1A2) and ArsR-RFP''' |
** Take only the time range (0-60min) | ** Take only the time range (0-60min) | ||
Latest revision as of 20:18, 30 September 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
GVP Cluster
Plates
| Name | Plasmid Used | Antibiotics on Plasmid | No. of Colonies | Date |
| Positive Control (GVP) | J61035 | Ampicillin | 11 | 28 sept. |
| Positive Control (GVP)(conc.) | J61035 | Ampicillin | ~130 | 28 sept. |
| GVP Ligation no.1 | J61035 | Ampicillin | 0 | 28 sept. |
| GVP Ligation no.1 (conc.) | J61035 | Ampicillin | 2 | 28 sept. |
| GVP Ligation no.2 | J61035 | Ampicillin | 0 | 28 sept. |
| GVP Ligation no.2 (conc.) | J61035 | Ampicillin | 3 | 28 sept. |
| Plate 1, 1M | pSB1A2 | Ampicillin | ~20 (red) | 28 sept. |
| Plate 1, 1M (conc.) | pSB1A2 | Ampicillin | ~200 (red) | 28 sept. |
| Plate 1, 2M | pSB1A2 | Ampicillin | ~25 | 28 sept. |
| Plate 1, 2M (conc.) | pSB1A2 | Ampicillin | ~150 | 28 sept. |
- → The few colonies on the GVP ligation plates might be incorrect ligations, as was the case before. Restriction analysis should confirm positive or negative results.
- → The red colonies on the plates of transformed E.coli DB3 cells with plasmids from the cup labeled "B0034", indicates it is not B0034, but the plasmid located one position to the left (1-1M) with RFP coding device. Unfortunately, this plasmid was transformed as being B0034, and was stored in the -80 fridge at position 19. For future work the cup will be removed and two new cups will be made (containing 1-1M and 1-2M).
The E.coli DB3 cells seem to be less competent than the previous E.coli TOP10 cells, and starting at the end of the week E.coli TOP10 will be used again.
New Cultures
From each plate two colonies were used to inoculate 6mL LB-amp100 medium, and the tubes were stored at 37C overnight in a waterbath.
Transporters
Metal Accumulation
- Redo As(III) uptake experiment for E. coli WT (pSB1A2) and ArsR-RFP
- Take only the time range (0-60min)
Vectors
Dry
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