How different concentrations of Running Buffers effects Gel Electrophoresis

AE and TBE Running Buffers WHAT ARE TRIS ACETATE EDTA AND TRIS BORATE EDTA? Tris acetate EDTA (TAE) and tris borate EDTA (TBE) are the two most common running buffers used in nucleic acid electrophoresis. As buffers, they have a fairly constant pH and are able to conduct electricity because of their concentration of hydrogen ions. These properties are necessary for gel electrophoresis during which proteins are separated by electric charge. PREPARING RUNNING BUFFERS TBE and TAE are most often mixed from their constituent parts into laboratory stock solutions. Both buffers can be purchased at the working concentration or in a powdered or concentrated format that is simply prepared via dilution. Refer to the recipes below to prepare TAE and TBE in common stock solution concentrations. Our buffer calculator can also help you find the correct dilution (or conversion) for various buffers from stock solution to a desired molarity and volume. VIDEO: TAE AND TBE BUFFERS FOR GEL ELECTROPHORESIS Review common TAE and TBE buffer solution recipes and learn which running buffer to choose for your nucleic acid gel electrophoresis application. Research technology specialist Chris Lemke mixes up stock solutions and provides helpful buffer selection tips. TAE Buffer 50x Stock Recipe 242 g tris base in double-distilled H2O 57.1 ml glacial acetic acid 100 ml 0.5 M EDTA solution (pH 8.0) Adjust volume to 1 L. 10x TAE Recipe For 1L of 10x solution, 48.5 g tris 11.4 mL glacial acetic acid 20 mL EDTA (pH 8.0) 1x TAE Recipe Dilute 1:10 0.4 M tris acetate (pH approximately 8.3) M EDTA using ultrapure water. TBE Buffer 10x Stock Recipe 108 g tris base 55 g boric acid 900 ml double-distilled H2O 40 ml 0.5 M EDTA solution (pH 8.0) Adjust volume to 1 L. 1x TBE Preparation Dilute 10x concentrated TBE buffer 10-fold with ultrapure water. The final solution should contain: M tris (pH 7.6) 45 mM boric acid 2.5 mM EDTA Buffer Prep Tips If precipitation is present, warm to 37 °C and mix until completely dissolved prior to dilution. It is recommended 1x working solutions be filtered through a 0.2 mm filter before use. 1x working solutions can be used until the expiration date on packaging with storage at room temperature. Discard if buffer becomes cloudy or discolored. #NikolaysGeneticsLessons #GelElectrophoresisBuffer #TrisBorateEDTABuffer #Borate #BoricAcid #EDTA #PCR #PolymeraseChainReaction #electrophoresis #trishydroxymethylaminomethane #THAM #Tris #EthylenediaminetetraaceticAcid #gelElectrophoresis #science #Education #equipment #Labs #SeedingLabs #Buffers #TeleScience #molecularBiology #DNA #Proteins #nucleicAcids #gelElectrophoresisExplained #gelElectrophoresisOfDna #electrophoresisOfDna