PCR from single colonies?

Robert Horton horton at biosci.cbs.umn.edu
Wed Jan 17 17:51:33 EST 1996


Jennifer Lyon (JLyon at novagen.com) wrote:
: Nathan Lange wrote:
: > 
: > Does anyone have a good protocol or advice for doing PCR directly from
: > single bacterial colonies?

: This is from my company's pET Manual: 
: 1. Pick a colony from an agar plate and transfer the bacteria to a 1.5ml 
: tube containing 50 ul of sterile water. Vortex.
: 2. Boil (or heat block at 99degrees) for five minutes
: 3. centrifuge at 12,000xg for 1 min
: 4. Transfer 10 ul of the supernatant to a fresh 0.5ml tube for PCR, leave 
: on ice until use.
: 5. Procede to PCR (50 ul reaction usually)
: Hope it helps.

Alternatively:
1. Pick a colony with a sterile toothpick.
2. Dip the tip into a 10ul PCR reaction.
3. Cycle.
Too many bugs will kill the reaction.

-Bob


---
Robert M. Horton
http://134.84.47.3                        Have a :) day




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