Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/05/05

From OpenWetWare

Jump to: navigation, search
Plasmid Verification (multiple) Main project page
Previous entry      Next entry

Overview

All six liquid cultures grew as expected. Harvested cells and performed a plasmid miniprep for each. The BioBrick cloning constructs ( from KAH57 & KAH104) I'm naming DBN006.

DNA Concentration

Sample ID Conc. (ng/µL) St Dev (ng/µL) 260/280
DBN006 1383.43.201.94
DBN006 2671.211.911.93
DBN006 3298.67.321.94
KAH57259.33.251.93
KAH104293.744.451.91
pMaxGFP549.57.521.92

Will verify via restriction digest tomorrow.

Restriction Digest (05/06)

Part Conc. (ng/µL) Volume for 600ng (µL) Volume water (µL)
DBN006 1383.41.623.4
DBN006 2671.20.924.1
DBN006 3298.62.023.0
KAH57259.32.322.7
KAH104293.72.023.0
Reagent Volume (µL)
DNA + water25
10x FD Buffer (Green)3
EcoRI1
SpeI1
Total30

Incubate at 37°C for 10 minutes, then run total volume on the gel.

Digest Results

Colonies 1 and 3 contain inserts of the expected size.


Personal tools