Purpose:
The purpose of this lab is to successfully isolate DNA from our cheek cells and to prepare a PCR reaction for the amplification of an Alu insert.
Materials:
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Procedure:
DNA Preparation
- Swirl 10 ml of saline in your mouth for 30 seconds.
- Spit into cup and swirl to mix cheek cells.
- Put 1000 ul of cheek cell solution into 1.5 ml microfuge tube.
- Microfuge tube.
- Pour off supernatant leaving 100 ul covering cell pellet.
- Put 50 ul of cell suspension into tube of chelex.
- Heat on heat block for 10 minutes.
- Release pressure out of tube and microfuge.
- Take 50 ul of supernatant from tube and place in new microfuge tube.
PCR
- Pipet 20 ul of master mix, 20 ul of primer mix, and 10 ul of DNA into new PCR tube.
- Place in thermal cycler.
Gel
- 50 ml of 1x TAE an 1 g agarose
- Heat solution until dissolved.
- Pour in mold when cool.
- 20 ml of DNA into new tube with 4 ml loading dye
- Centrifuge and load 20 ul into well.
- Record lane and gel number