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When thinking about the percentage of agarose to use in the gel, it is important to understand your resolution needs. What this means is that the concentration of agarose should be able to separate your desired fragment size enough so that it is identifiable. Take the example figure below of an artificial agarose gel comparison between a % and % gel. Notice that a % gel did not separate (resolve) the two bands at 400 bp and 500 bp that well, and the last bands did not separate at all (100 bp and 60 bp). Now when running the same samples on a % gel, the resolution was increased whereby these smaller bands are now seen as distinct products. To help decide what percentage of agarose to use, I have included a reference table below containing different agarose gel concentrations, along with the corresponding resolution sizes (adapted from Thermo Fisher). To use the table, look at the Resolution column and identify the range which includes your molecular size of interest. For example, if I want to separate a 100 bp product from a 1,500 product on a gel, I would select the % agarose gel since the resolution is between 50 – 2,000 bp. Agarose % (w/v) Resolution % 1,000 – 30,000 bp % 800 – 12,000 bp % 500 – 10,000 bp % 400 – 7,000 bp % 200 – 3,000 bp % 50 – 2,000 bp If there are a wide range of sizes to be separated on a gel, it is recommended to start with a % agarose gel concentration. #NikolaysGeneticsLessons #gelElectrophoresis #gelElectrophoresisExplained #gelElectrophoresisOfDna #gelElectrophoresisProcedure #gelElectrophoresisLab #gelElectrophoresisAnalysis #agaroseGelElectrophoresis #agaroseGelElectrophoresisProtocol #agaroseGelElectrophoresisOfDna #agaroseGelElectrophoresisExplained #dnaGelElectrophoresis #dnaElectrophoresis #electrophoresisOfDna #sdsPageGelElectrophoresis #polyacrylamideGelElectrophoresis #glycerol #TBE
