Oct 18, 2019

Public workspacecolony PCR

DOI
dx.doi.org/10.17504/protocols.io.79qhr5w
  • 1Heinrich-Heine Universität Düsseldorf
Igem Dusseldorf
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DOI: dx.doi.org/10.17504/protocols.io.79qhr5w
Protocol Citation: Igem Dusseldorf 2019. colony PCR. protocols.io https://dx.doi.org/10.17504/protocols.io.79qhr5w
Manuscript citation:
https://www.promega.com/-/media/files/resources/protocols/product-information-sheets/g/gotaq-green-master-mix-protocol.pdf
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
We use this protocol and it's working
Created: October 15, 2019
Last Modified: October 18, 2019
Protocol Integer ID: 28688
Abstract
colony PCR suitable for Synechocystis
Materials
2X GoGreen PCR Mix (or other MasterMix with e.g. redtaq also possible)
MilliQ
forward and reversed Primer
Start
Start
transfer some cells to 10 µL sterile MilliQ
heat 98°C
Duration00:07:00

use 1 µl of reaction for colony PCR
Protocol
Protocol
you can also use redtaq polymerase etc.
componentsamount per reaction [µl]
2X GoGreen PCR Mix5
fwd Primer1
rev Primer1
MilliQ3


Temperature[°C]Time [s]
94180
9430Repeated 30 times
variable, depends on annealing temp. of primer30
72130
72420

2 µl of reaction can be used for loading the gel