Team:Groningen/Template/MODULE/Notebook/toolbox/week10

From 2014.igem.org

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For each transformation, two seperate cultures were grown in LB with 10 &mu;g/ml
For each transformation, two seperate cultures were grown in LB with 10 &mu;g/ml
chloramphenicol. A miniprep was done on the cultures and the insert size
chloramphenicol. A miniprep was done on the cultures and the insert size
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was checked with colony PCR with VF2 (BBa_G00100) and VR (BBa_G00101)
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was checked with colony PCR, using <a href="http://parts.igem.org/Part:BBa_G00100">VF2</a> and <a href="http://parts.igem.org/Part:BBa_G00101">VR</a> as
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primers and the a restriction analysis was done using EcoRI and PstI.
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primers. In addition, a restriction analysis was done using EcoRI and PstI.
Cultures that showed a correct insert size were sent for sequencing.
Cultures that showed a correct insert size were sent for sequencing.
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Latest revision as of 01:21, 18 October 2014

14 September - 21 September
 
Because of the constant low concentrations of the plasmid pSB1C3 with the new BioBricks, it was decided to transform the isolated pSB1C3 with the new BioBricks (NisA, NisC, NisRK, PNisI and sfGFP(Bs)) again in Escherichia coli DH5α. The pSB1C3 plasmids containing the new BioBricks were again isolated. For each transformation, two seperate cultures were grown in LB with 10 μg/ml chloramphenicol. A miniprep was done on the cultures and the insert size was checked with colony PCR, using VF2 and VR as primers. In addition, a restriction analysis was done using EcoRI and PstI. Cultures that showed a correct insert size were sent for sequencing.