"
Team:Evry/Notebook/July/4
From 2012.igem.org
(Difference between revisions)
(Created page with "What we did today: <br> <br> 1) Gel electrophoresis of DNA digests (from 28.06.)<br> followed by cutting-of bands of interests (pCS2+ plasmid and XFP plasmids) <br> 2) Gel-extrac...") |
|||
| Line 13: | Line 13: | ||
<br> | <br> | ||
Incubation 3h at 37 degrees | Incubation 3h at 37 degrees | ||
| + | <html> | ||
| + | |||
| + | <center> | ||
| + | <a href="https://2012.igem.org/Team:Evry/Notebook/July/3"><img src="https://static.igem.org/mediawiki/2012/e/ed/Previous_day_arrow.png"></a> | ||
| + | |||
| + | <a href="https://2012.igem.org/Team:Evry/Notebook/July/5"><img src="https://static.igem.org/mediawiki/2012/e/e3/Next_day_arrow.png"></a> | ||
| + | </center> | ||
| + | </html> | ||
Revision as of 16:27, 5 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
