Jul 12, 2022

Public workspaceLAB Agarose Gel Electrophoresis Buffer Recipe

DOI
dx.doi.org/10.17504/protocols.io.e6nvwkyedvmk/v1
  • 1University of Wisconsin - Stout
  • Yeast ORFans CURE
Brian Teague
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DOI: dx.doi.org/10.17504/protocols.io.e6nvwkyedvmk/v1
External link: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0011318
Protocol Citation: Brian Teague 2022. LAB Agarose Gel Electrophoresis Buffer Recipe. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwkyedvmk/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 in an introductory molecular cell biology lab course and it's working.
Created: July 12, 2022
Last Modified: July 12, 2022
Protocol Integer ID: 66462
Keywords: agarose, gel, electrophoresis, lithium, acetate
Abstract
A recipe to make lithium acetate / borate agarose gel electrophoresis buffer. This buffer has a lower ionic strength than TAE, but maintains its ability to resolve both small and large bands.

Additionally, it is MUCH less expensive, and if you are short on time (like in a lab course!) you can run the gel at a higher voltage.

From https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0011318 via https://bitesizebio.com/25078/faster-even-cooler-dna-gels/
Image Attribution
By TransControl - english wikipedia, CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=2046526
Guidelines
Use good laboratory practices for measuring weights and volumes.
Materials
ReagentLithium Acetate DihydrateSigma AldrichCatalog #L4158
ReagentBoric acidFisher ScientificCatalog #BP1681
ReagentPaper pH StripsThermo Fisher ScientificCatalog #13-640-508

Protocol materials
ReagentBoric acidFisher ScientificCatalog #BP1681
ReagentPaper pH StripsThermo Fisher ScientificCatalog #13-640-508
ReagentLithium Acetate DihydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #L4158
ReagentLithium Acetate DihydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #L4158
ReagentBoric acidFisher ScientificCatalog #BP1681
ReagentPaper pH StripsThermo Fisher ScientificCatalog #13-640-508
Safety warnings
Lithium acetate: May cause eye and skin irritation. May cause respiratory and digestive tract irritation. The toxicological properties of this material have not been fully investigated.

Boric acid: May damage fertility. May damage the unborn child.

Wear appropriate personal protective equipment (PPE), including a lab coat, nitrile gloves and safety glasses.

25X Buffer Concentrate
25X Buffer Concentrate
Weigh out Amount25.5 g ReagentLithium Acetate DihydrateSigma AldrichCatalog #L4158

Note
Make sure it's the dihydrate, not anhydrous lithium acetate! If you're using anhydrous, weigh out Amount16.5 g instead.


Weigh out Amount15.5 g ReagentBoric acidFisher ScientificCatalog #BP1681 .

Mix the lithium acetate and boric acid with deionized water to a final volume of Amount1 L

Pipette a small amount onto a ReagentPaper pH StripsThermo Fisher ScientificCatalog #13-640-508 . Make sure the pH is between 6.5 and 7.0.

1X Buffer
1X Buffer
Dilute Amount40 mL of 25X stock solution with Amount960 mL deionized H2O. Mix well.