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Lab 10-Gel - – IT IS A CARCINOGEN 7 Remove comb place...

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Lab 10 Electrophoresis 50X TAE electrophoresis buffer: 242g Tris Base 57.1ml Glacial Acetic Acid 100ml 0.5M EDTA (pH 8) 1L total Procedure: 1. Make 1L of 1X TAE buffer (provided). 2. Mix 1g agarose with 100ml 1X TAE buffer in a 125ml Erlenmeyer flask. Cover with plastic wrap. 3. Microwave 30seconds, stir, and microwave again 30 seconds, or until dissolved. 4. Cool on a hot plate until flask is “touchable”. 5. Prep the gel apparatus: use black ‘dam’ in the gel tray and insert the proper comb. 6. Add 0.5 μ l Ethidium Bromide to bottom of gel tray. Pour agarose into tray and allow to harden. USE EXTREME CAUTION WHEN USING ETHIDIUM BROMIDE
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Unformatted text preview: – IT IS A CARCINOGEN!!! 7. Remove comb, place tray into gel apparatus and fill with 1X TAE buffer until gel is covered. 8. Pipet 10ul of DNA ladder into the first lane (prepared for you), and follow suit with PCR sample in remaining lanes. 9. Put on apparatus cover and run gel at 100V for 30 minutes. 10. Using the imaging apparatus, take a picture of the gel under UV light. MAKE SURE THE UV LIGHT IS COVERED- do not expose eyes to UV light! 11. Compare your PCR product with the 1Kb ladder to determine the size of the band....
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