Jan 04, 2016
Colony PCR Protocol
- Matthew Sullivan1
- 1Matthew Sullivan Lab, University of Arizona, Ohio State University
- VERVE Net
- Sullivan Lab
Protocol Citation: Matthew Sullivan 2016. Colony PCR Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.c6czav
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: June 22, 2015
Last Modified: September 14, 2018
Protocol Integer ID: 932
Guidelines
Note: complete run takes about 5 hours, most of which does not require your direct attention.
Ligated DNA is transformed into cells
Plate transformed cells on selection plate
Go home, wait 13 hours
13:00:00
Pick a well separated single colony from selection plate via yellow pipette tips
Move said colony to 96 well microtittle plate
Place individual colony into a single well containing 50 µl sterile distilled water
Repeat for as many colonies as desired
Make up PCR mix
Use multichannel pipette to add 7.5 µl PCR mix to a 96 well PCR plate
Add 2.5 µl colony solution
Run PCR reaction
Note
Do something else while waiting.
Add 4 µl loading dye once PCR run is complete
Load (PCR reaction mix + dye) into gel and run
Complete run takes about 5 hours
05:00:00