Oct 21, 2021

Public workspaceAgarose gel electrophoresis V.2

This protocol is a draft, published without a DOI.
  • Yuichiroh Ikagawa1
  • 1iGEM Gifu
  • iGEM Gifu
Yuichiroh Ikagawa
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Protocol Citation: Yuichiroh Ikagawa 2021. Agarose gel electrophoresis. protocols.io https://protocols.io/view/agarose-gel-electrophoresis-bzcwp2xeVersion created by Yuichiroh Ikagawa
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: October 21, 2021
Last Modified: October 21, 2021
Protocol Integer ID: 54390
Keywords: agarose gel electrophoresis electrophoresi, gel electrophoresi, identifying dna, agarose gel, dna, dna by molecular weight, restriction enzyme treatment, pcr
Abstract
Electrophoresis is the most basic technique for identifying DNA amplified by PCR or restriction enzyme treatment. Agarose gels are used to separate the DNA by molecular weight. Here we describe the technique using a 1.5 % agarose gel.
Materials
Reagents
Agarose S agarose powder (Nippon Gene)
x50 TAE buffer
DNA sample
x6 DNA loading dye (New England Biolabs)

Equipment
Mupid® exU electrophoresis machine (Mupid)


Troubleshooting
Weigh out 0.75 g of agarose powder and add it to the triangular flask.
Add 49 ml of sterile water to the first flask and shake gently.
Add 1 ml x50 TAE buffer to the flask and heat in the microwave.
Allow the melted agarose to cool for 5 minutes before pouring it into a template set with a comb.
Cover with plastic wrap and leave for 15 minutes until the gel has solidified.
Mix 1 ul of loading dye with 5 ul of sample DNA.
If necessary, the sample DNA solution can be diluted and added.
Carefully apply 6 ul of the DNA-dye mixture to the wells.
Run the electrophoresis 100 volts for 30 minutes.
Remove the gel from the electrophoresis bath and stain with ethidium bromide (EtBr) for 20 minutes.
Check the stained DNA with a transilluminator.