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From 2012.igem.org

I. AGE of PCR Samples

• Make Gel:

• Measure out 0.5 g of agarose and add to a 250 mL E-flask
• Add 50 mL of 0.5x TBE buffer and swirl to mix
• Cover flask with a Kimwipe and microwave for ~1 minute until clear
• Allow to cool to ~60 ^oC and pour into beaker for ethidium bromide (EtBr)
• Add ~1.5 uL of EtBr stock to agarose and swirl to mix
• Immediately pour gel into tray with combs
• Allow to solidify, then remove the comb and tape and place the tray in the gel box with the wells closer to the black electrode

• Make sure it is submerged in TBE buffer

• Load Gel:

• Mix samples as drops on a piece of parafilm

• Mix:

Ladder (Invtrogen 1 kb plus)	Blank	Sample
*5 uL DNA ladder *1 uL 6x Loading Dye	*8.3 uL dH2O *1.7 uL 6x Loading Dye	*6.3 uL dH2O *1.7 uL 6x Loading Dye *2.0 uL DNA

• Load ~6 uL on gel according to the following chart:

Lane 1	2	3	4	5	6	7	8
NEB 2-log ladder	Blank	-	+ 1	+ 2	+ 3	+ 4	+ 5

• Store DNA at 4 ^oC

• Run Gel:

• Close gel box and turn on power pack
• Run gel at 30 V for 20 min to stack gel
• Run gel at 75 V until the markers have reached ~3/4 down the gel

• Image Gel:

• RESULTS:

Source: Media:Srk_2012-07-28_18hr_04min.tif

Feature	Expected Size (bp)
Cassette	5152