Jun 26, 2023
Genotyping protocol for mice
- vanessa promes1
- 1Northwestern University, Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815
Protocol Citation: vanessa promes 2023. Genotyping protocol for mice. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx3mjwg8j/v1
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: June 26, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 84056
Keywords: ASAPCRN, Genotyping, mice
Funders Acknowledgement:
Aligning Science Across Parkinson's [ASAP-020600] through the Michael J. Fox Foundation for Parkinson's Research (MJFF)
Grant ID: ASAP-020600
Abstract
This protocol describes the general method to genotype the mice.
DNA Extraction
DNA Extraction
30m
30m
Obtain tail sample from mice.
Add 100 µL of NaOH into tail sample and place in hot plate at 100C for 00:10:00 . Vortex sample and place in hot plate once more for 00:10:00 .
20m
Add 50 µL of 1M Tris pH 8.0 to neutralize extraction
Spin for 20,000xg for 00:10:00
10m
Use 2µL in PCR reaction.
Preparing PCR reaction
Preparing PCR reaction
Obtain a 1.5ml microcentrifuge tube and add 10ul 2x DreamTaq Green Master mix. 2ul forward and reverse primers, 2µL DNA lysate and Nuclease-free water up to 20µL
Thermocycling parameters:
95oC 5min
95oC 30sec| x 30 cycles
60oC 30sec|
72oC 1min|
72oC 2min
4oC hold
Run DNA Sample
Run DNA Sample
In 2% agarose in TAE buffer add 10ul of the sample and run for around 30min