Mar 12, 2023

Ai14 Genotyping 

DOI
https://dx.doi.org/10.17504/protocols.io.81wgbywy1vpk/v1
  • 1BYU
Ronald.zegarra
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DOI: https://dx.doi.org/10.17504/protocols.io.81wgbywy1vpk/v1
Protocol Citation: Ronald.zegarra 2023. Ai14 Genotyping . protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbywy1vpk/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: In development
We are still developing and optimizing this protocol
Created: March 10, 2023
Last Modified: March 12, 2023
Protocol  Integer ID: 78457
Keywords: ai14
Abstract
genotyping
Thaw reagent on ice for00:10:00 minutes

10m
PrepareMaster Mix :

Required
13.1 uL milliQ
Required
2 uL 10x PCR Buffer
Required
0.5 Advance 2 Polymerase Mix
Required
0.4 uL 10 mM dNTP
Required
1 uL Ai14 WT Forward
Required
1 uL Ai14 WT Reverse
Required
1 uL Ai14 Mutant Forward
Required
1 uL Ai14 Mutan Reverse

Pipette 19 µL of Master Mix into 0.2 mL Tube


Lightly Vortex DNA sample for 00:00:30

30s
Add 1 µL of DNA to each tube

Select "Ai14" PCR protocol on the thermocycler
Run 15 uL of PCR samples and a DNA ladder on a
Required
200 uL 1x TAE
Required
4 mg of agarose
at 120 V for 01:00:00 or until the dye is somewhat in the middle of the gel.

1h