Luckau Protocols

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Contents

Tara Luckau Protocols

Clark Laboratory

Agarose Gel

Purpose

Agarose gels are used to verify the presence and estimate the size of DNA (both genomic and amplified). When loaded into a gel and subjected to an electric current, DNA fragments migrate to the positive terminal. Small DNA fragments migrate more rapidly than large ones. By running a commercially available ladder of fragments of known size, we can estimate the DNA fragment size in our sample of interest.

To visualize the DNA after electrophoresis, a dye called Gel Red is added during the agarose-making process. The dye binds to the DNA and flouresces when exposed to UV light.

Materials To Be Familiar With

  • Gel Rig, caster tray, combs
  • Image:OwlA2Large.jpg

Protocol

  1. Make gel using 1x TAE and agarose
    • Use 1% for genomic DNA, 2% for amplified DNA
Gel Rig Size 1x TAE (mL) 1.5% agarose (g) 2% agarose (g) Gel Red (µL) Sample volume (µL) max voltage (V)
small 50 _ 1 5 4-6 80
medium 130 _ 2.6 13 24-tooth comb: 6-10 120
36-tooth comb: 3-5
large _ _ _ _ _ 170
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