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From 2012.igem.org

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Latest revision as of 15:25, 12 July 2012

What we did today:

1) Gel electrophoresis of DNA digests (from 28.06.)
followed by cutting-of bands of interests (pCS2+ plasmid and XFP plasmids)
2) Gel-extraction (Kit used: QIA quick gel extraction) followed by nano-drop measurements in order to get information about DNA concentration.
Result: very low quantity of DNA. We realized that quantity of DNA used for digestion was too low (less than 2 micrograms of DNA/sample).
3) DNA Digestion (of pCS2+, CFP, RFP, GFP, YFP). Protocol:

Final volume of sample: 30 uL

 3 uL Buffer 10x (choose a right buffer for each enzyme you use)

 1 uL Restriction Enzyme 1 

 1 uL Restriction Enzyme 2 

 2 ug DNA (calculate the volume which you need to add, depends of the DNA concentration) 

 c uL ddH2O (fill till 30 uL) 

Incubation 3h at 37 degrees