Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2016/02/08
From OpenWetWare
Today's project is... | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||
OverviewMore adventures in cloning... To do:
DNA Quantificationquantification went fine, concentrations are between 50 and 70 ng/µL for DBN010. A bit lower for HA1_v0120, between 7 and 24 ng/µL. 260/280 values are good. Restriction Digest
Incubate at 37 °C for 10 minutes, then purify via gel electrophoresis. Gel purificationAll four plasmids appear to contain the insert. Will proceed with gel extraction from colonies 1 and 3. DNA Quantification, post-purificationConcentration is ~7 ng/µL for both samples. 260/280 is on the high side, around 2.8-2.9. Going to proceed with ligation... Ligation
Incubate at room temperature for 10 minutes, then take 1 µL from each sample (treatment & negative control) and use for quick transformation protocol.
|