Oct 24, 2019

Public workspacePCR V.2

DOI
dx.doi.org/10.17504/protocols.io.8nyhvfw
  • 1Universidad Complutense de Madrid
  • AEGIS - Madrid iGEM 2019
Laura Armero
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DOI: dx.doi.org/10.17504/protocols.io.8nyhvfw
Protocol Citation: Claudia Troncone Clemente 2019. PCR. protocols.io https://dx.doi.org/10.17504/protocols.io.8nyhvfw
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 24, 2019
Last Modified: October 24, 2019
Protocol Integer ID: 29112
Abstract
Our aim with this protocol is to amplify DNA. This protocol has been optimized has a general amplification

As the quantity of DNA is exponentially increased during the performance of the selection, further modification in the numbers of cycle will be needed to be implemented.
Guidelines
We sure to have all the surfaces and materials clean before the start.
All the procedures must be done in an sterile environment to avoid contamination.
Materials
MATERIALS
ReagentSpeedy Supreme Green Master MixNZYTechCatalog #MB39102
ReagentAgarose (LM-ultrapure grade)NZYTechCatalog #MB123
  • Aptamer library (order to IDT)

  • Forward primer (order to IDT):

  • Reserve primer (order to IDT):

  • Thermocycler
  • TAE buffer
Prepare the PCR reaction mixture following the specifications below:



Perform the amplification in a general thermocycler in the following conditions. Adjust the annealing temperature according to the primers used, and the hotstart to the specifications of your polymerase:



Prepare a 3% agarose gel. Load the samples and perform the electrophoresis at 90V for 50 min.
Remove the gel and observe the bands under UV light.