Microwave for approximately 1 min or until the agarose is fully dissolved
Once the solution has cooled to the touch, add 2.5 uL of Smart Glow 20,000X Gel Stain
Pour the solution onto a gel tray with a comb in it
Once it has solidified, put the gel tray into a gel box and cover the gel with TAE. Remove the comb.
Mix samples with 5X loading dye. Load the wells with NEB 1 kb plus ladder and the samples.
Run at 120-140 V until the dye is about 2/3 of the way down the gel.
View under blue light reader.
Gel Extraction (with QIAquick Kit from Qiagen)
Excise the band of DNA with a razor blade. Trim as much gel as possible and have the blue light on for a minimal amount of time.
Weigh the gel slice. Add 3 volumes of Buffer QG to the tube containing the slice.
Incubate in a water bath at 50 C until the gel slice has completely dissolved. Vortex every 2-3 minutes to help it dissolve. The color of the solution should be yellow.
Add 1 volume of isopropanol to the mix.
Add sample to a QIAquick spin column inside a 2 mL collection tube and centrifuge for 1 min at 17,900 rcf (13,000 rpm).
Discard flow-through. Add another 500 uL of Buffer QG to the column and centrifuge for another 1 min.
Discard flow-through. Add 750 uL of Buffer PE to the column and spin for 1 min.
Discard flow-through. Centrifuge for an additional 1 min to remove residual ethanol.
Transfer column to a fresh 1.5 mL microcentrifuge tube. Add 30 uL of Elution Buffer to the center of the membrane and let sit for 5 mins.
