Assembly Check of PCR Reactions 1,2,3,4
Follow steps for gel electrophoresis.
- Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray.
- Create 1% gel by putting .6 grams of agarose into flask.
- Microwave agarose solution for 30 seconds
- Using hot gloves, grab flask out of microwave and swirl flask vigorously. Place back in microwave for 30 more seconds.
- When flask is taken out of microwave, make sure that the agarose is completed dissolved.
- Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the thin "teeth" and slip in the notches.
- Add EtBr (ethidium bromide) to the agarose gel. There should be 1 microliter of EtBr per 10ml of agarose gel. Therefore, 6 µL.
- Pour gel into tray. Wait twenty minutes for gel to settle.
- Wash the agarose gel flask.
- Pipette 15 μL of DNA ladder into first well. Pipette 5μL of PCR reactions into subsequent wells.
- Turn on electrophoresis and let run for thirty minutes.
The assembly did not work. No bands were seen when placed under UV light.