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Team:Groningen/Notebook/20 July 2009
From 2009.igem.org
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Wet
GVP Cluster
Precipitation of the restriction fragments from GVP, J23109, J23100 and J23106
To concentrate BBa_J23109, BBa_J23100, BBa_J23106 and GVP it is first precipitated and then MQ is added.
Procedure:
- 100 μl absolute ethanol is added to ~50 μl Restriction fragment of GVP (12.1 ng/μL),J23109 (13.1 ng/μl), J23100 (11.9 ng/μl) and J23106 (11.1 ng/μl).
- Incubation -80°C for 1 h
- Centrifugation 30 min 0°C (14000rpm)
- Supernatant is removed
- Wash with 1000 μl 96% Ethanol (added ethanol and inverted a couple of times)
- Centrifugation 10 min 4°C (14000rpm)
- o/n air dried on bench top
Preparation of 1-14N and 1-1D for glycerol stocks, restiction analysis and 3A assembly
1-14N () and 1-1D () are inducible promoters, induced by L-arabinose and lactose (or IPTG) respectively.
- DNA from iGEM plates resuspended in 15 μL MQ and stored at -20°C
Transporters
Below the PCR of 17/07, There seems to be a vague band at ~1150 in lane 1.
The band at ~1150 kb was cut out of the gel and used for pcr described below.
Metal AccumulationVectorsPositive control for the psB1AC3 vector done with both new and last years primers.
The gel shown below shows bands at about 1000 which is not what would be expected. Band of 315 were expected for lanes 1-6.
Dry
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