Tara Luckau Protocols
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
- 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)|
|medium||130||_||2.6||13||24-tooth comb: 6-10||120|
|36-tooth comb: 3-5|