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Team:Groningen/Notebook/18 August 2009
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===Metal Accumulation=== | ===Metal Accumulation=== | ||
| - | {{ | + | {{done| Restriction analysis MBP-ArsR}} |
| - | + | <center>[[Image:MBP-ArsR_Fusion_protein_in_pSB1AC3_restricted_with_XbaI_%26_EcoRI_-_18august2009.jpg| 500px]]</center><br> | |
| + | 1% Agarose (1xTBE) gel with restriction analysis of MBP-ArsR fusion protein plasmids, with selfcloser (1), sample (white colony) (2), sample (red colony??) (3). Expected sizes of positive construct are shown. Samples were digested using [http://www.fermentas.com/catalog/re/fastpsti.htm PstI] & [http://www.fermentas.com/catalog/re/fastxbai.htm XbaI], ~45 min 37 °C. | ||
| + | Sample MBP-ArsR (200) was isolated from red organims, indicating the presence of RFP or similar, this was however not engineered in this plasmid. Therefore a deviant restriction pattern was to be expected. It appears as if two different plasmids were present in this sample. | ||
| + | |||
===Vectors=== | ===Vectors=== | ||
{{todo| Restriction analysis metal promotors}} | {{todo| Restriction analysis metal promotors}} | ||
Revision as of 13:49, 18 August 2009
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Wet
GVP Cluster
Transporters
HmtA curios about the dirty experiment. a 1% gel control will show if expected product is there. Unfortunately there was no product, the primer dimer changed and pcr 1 and 2 remain available. PCR again. Different program. Plus starting an other PCR1 and PCR2 to get new megaprimers.
Metal Accumulation
Restriction analysis MBP-ArsR
1% Agarose (1xTBE) gel with restriction analysis of MBP-ArsR fusion protein plasmids, with selfcloser (1), sample (white colony) (2), sample (red colony??) (3). Expected sizes of positive construct are shown. Samples were digested using PstI & XbaI, ~45 min 37 °C. Sample MBP-ArsR (200) was isolated from red organims, indicating the presence of RFP or similar, this was however not engineered in this plasmid. Therefore a deviant restriction pattern was to be expected. It appears as if two different plasmids were present in this sample.
Vectors
Restriction analysis metal promotors
Dry
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