Purpose:
The purpose of the lab is to isolate our cheek cells and examine if they contain the Alu repeat in one part of our chromosomes.
Materials:
9% Saline solution
Micropippettes + tips
Waste container
Microcentriruge + racks
Microcentrifuge tubes
PCR tubes
Agarose + 1X TAE
Gel chambers + molds
Load dye
Primer mix
Master mix
Chelex solution
Water
Positive control DNA
Procedure:
I. DNA Preparation
1. Swirl saline solution in mouth, spit and swirl in cup
2. Put solution into tube and centrifuge
3. Pour out excess solution so its mostly just cells left
4. Resuspend cells and put into Chelex solutions
5. Heat solution for 10 mins and then centrifuge again
6. Insert "Isolated DNA" into new tube
II. PCR
1. Put 20 ul of Master and Primer mix into a PCR tube
2. Add 10 ul of the DNA
3. Set up 2 tubes of each control
4. Place each tube in a thermal cycler
III. Make Gels
1. Cmbine 50 ml of 1X TAE and 1 gram of Agarose
2. Heat until fully dissolved
3. Pour into gel chamber
IV. Gel Electrophoresis
1. Spin DNA
2. Take 20 ul of DNA and put into 1.5ml tube
3. Put 4 ul of dye into tube
4. Spin them together in centrifuge
5. Insert 20 ul into gel chambers
The purpose of the lab is to isolate our cheek cells and examine if they contain the Alu repeat in one part of our chromosomes.
Materials:
9% Saline solution
Micropippettes + tips
Waste container
Microcentriruge + racks
Microcentrifuge tubes
PCR tubes
Agarose + 1X TAE
Gel chambers + molds
Load dye
Primer mix
Master mix
Chelex solution
Water
Positive control DNA
Procedure:
I. DNA Preparation
1. Swirl saline solution in mouth, spit and swirl in cup
2. Put solution into tube and centrifuge
3. Pour out excess solution so its mostly just cells left
4. Resuspend cells and put into Chelex solutions
5. Heat solution for 10 mins and then centrifuge again
6. Insert "Isolated DNA" into new tube
II. PCR
1. Put 20 ul of Master and Primer mix into a PCR tube
2. Add 10 ul of the DNA
3. Set up 2 tubes of each control
4. Place each tube in a thermal cycler
III. Make Gels
1. Cmbine 50 ml of 1X TAE and 1 gram of Agarose
2. Heat until fully dissolved
3. Pour into gel chamber
IV. Gel Electrophoresis
1. Spin DNA
2. Take 20 ul of DNA and put into 1.5ml tube
3. Put 4 ul of dye into tube
4. Spin them together in centrifuge
5. Insert 20 ul into gel chambers