Mar 24, 2018
- Mauricio Durigan1
- 1State University of Campinas
External link: https://doi.org/10.1371/journal.pntd.0006928
Protocol Citation: Mauricio Durigan 2018. TOUCHDOWN PCR. protocols.io https://dx.doi.org/10.17504/protocols.io.nyzdfx6
Manuscript citation:
Durigan M, Cardoso-Silva CB, Ciampi-Guillardi M, Toledo-Silva G, Mori GM, Franco RMB, Souza AP (2018) Molecular genotyping, diversity studies and high-resolution molecular markers unveiled by microsatellites in Giardia duodenalis. PLoS Negl Trop Dis 12(11): e0006928. doi: 10.1371/journal.pntd.0006928
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: March 22, 2018
Last Modified: March 28, 2018
Protocol Integer ID: 11001
Keywords: protocol for the touchdown pcr, touchdown pcr, pcr assay, pcr, assay, protocol
Abstract
This is the protocol for the touchdown PCR assay
Troubleshooting
Before start
The assay has been optimized and validated for the Bio-rad C1000 Touch Thermocycler.
Cycling Parameters
94 oC 2 min
94 oC 1 min
65 oC ( -1 oC per cycle) 1 min
72 oC 2 min
94 oC 1 min
65 oC (-1 oC per cycle) 1 min
72 oC 2 min
Go to step 2, 9 times
94 oC 1 min
55 oC 1 min
72 oC 2 min
Go to step 9, 17 times
72 oC 7 min
15 oC FOREVER
End
Adapted from: Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS. “Touchdown” PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res. 1991;19: 4008. doi:10.1093/nar/19.14.4008