Team:Groningen/Notebook/22 July 2009
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- | As a check-up we also did a restrition analysis for the sPB3K3 vector with EcoR1 and Spe1 | + | As a check-up we also did a restrition analysis for the <partinfo>sPB3K3</partinfo>vector with EcoR1 and Spe1 |
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::* incubate at 37° for 30 min. | ::* incubate at 37° for 30 min. | ||
- | Results of the PCR and restiction shown below. | + | Results of the PCR and restiction shown below. Bands were expected in the pSB3K3 lanes with VR-VF2 primers at 316 kbp. |
[[image:F102471_2009-07-22_07hr_04min_ControlVRVF2_noted.jpg]] | [[image:F102471_2009-07-22_07hr_04min_ControlVRVF2_noted.jpg]] |
Revision as of 14:53, 22 July 2009
Wet
GVP Cluster
Transporters
Metal Accumulation
Vectors
In order to check the VR VF2 primers that were ordered this year a gradient PCR was done with a vector that is expected to work , a pUC vector and a negative control with no vector.
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As a check-up we also did a restrition analysis for the vector with EcoR1 and Spe1
10x fast digest buffer | 2 uL |
pSB3K3 (22,5 ngr/uL) | 8 uL (=180ngr up to 1ugr is recommended at fermentas) |
EcoR1 | 1 uL |
Spe1 | 1 uL |
MilliQ | 8 uL |
Total | 20 uL |
- Mix gently and spin down
- incubate at 37° for 30 min.
Results of the PCR and restiction shown below. Bands were expected in the pSB3K3 lanes with VR-VF2 primers at 316 kbp.
Dry
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