Oct 24, 2020
- 1Northeast Forest University
- 2020 iGEM NEFU China
Protocol Citation: Zhujun Wei 2020. PCR. protocols.io https://dx.doi.org/10.17504/protocols.io.bnwqmfdw
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, 2020
Last Modified: October 24, 2020
Protocol Integer ID: 43696
Materials
2×high Taq Master Mix
ddH2O
Template
F/R Primer
Bio-rad S1000TM Thermo Cycler.
Safety warnings
Please wear gloves for the experiment, don't try to touch the lid after PCR program initiation.
Before start
Set up a small box with ice, put DNA and 2×high Taq Master Mix into it before going into the Bio-rad S1000TM thermocycler cycler.Set up a small box with ice, put DNA and 2×high Taq Master Mix into it before going into the Bio-rad S1000TM Thermo Cycler.
Program the thermocycler as follows:
| Temperature | Time | |
| 98℃ | 5 min | |
| 98℃ | 15 s | |
| Tm-4℃ | 15 s | |
| 72℃ | (5-10s)× (1 kb) | |
| 72℃ | 5 min | |
| 4℃ | ∞ |
Add the following reagent to a PCR tube.(50 μl).
| 2×high Taq Master Mix (Enzyme) | 25 μl | |
| Template | 2 μl | |
| Forward Primer (10 μM) | 2 μl | |
| Reverse Primer (10 μM) | 2 μl | |
| ddH2O | 19 μl |