Alu PCR Lab
Objective:
To determine the frequency of the Alu insert among 5th period Bio/Biotech students.
Materials/Reagents:
1. 0.9% Saline Solution
2. Student Cheek Cells
3. 5% Chelex
4. Primer Mix Data:
5. Mater Mix (Nucleotides, DNA Polymerase, Buffer) (++) is 715bp
6. TBE Buffer (+ -) is 415bp,715bp
7. Agarose Gel, Tracking Dye (- -) is 415bp
8. DNA Stain "Gel Red" My results: (++)
9. Positive Control DNA
10. Base Pair Ladder
Procedure:
DNA Preparation:
1. Swirl 10 ml of saline in your mouth for 30 seconds.
2. Spit into cup and swirl to mix cheek cells.
3. Put 1000 ul of cheek cell solution into 1.5 ml microfuge tube.
4. Microfuge tube.
5. Pour off supernatant leaving 100 ul covering cell pellet.
6. Put 50 ul of cell suspension into tube of chelex.
7. Heat on heat block for 10 minutes.
8. Release pressure out of tube and microfuge.
9. Take 50 ul of supernatant from tube and place in new microfuge tube.
PCR:
1. Pipet 20 ul of master mix, 20 ul of primer mix, and 10 ul of DNA into new PCR tube.
2. Place in thermal cycler.
Gel:
1. 50 ml of 1x TAE an 1 g agarose
2. Heat solution until dissolved.
3. Pour in mold when cool.
4. 20 ml of DNA into new tube with 4 ml loading dye
5. Centrifuge and load 20 ul into well.
6. Record lane and gel number
Result:
My personal result was (++), meaning that both of my parents carry the Alu gene. After my results, I wonder if my 2 siblings also carry the Alu gene.
Objective:
To determine the frequency of the Alu insert among 5th period Bio/Biotech students.
Materials/Reagents:
1. 0.9% Saline Solution
2. Student Cheek Cells
3. 5% Chelex
4. Primer Mix Data:
5. Mater Mix (Nucleotides, DNA Polymerase, Buffer) (++) is 715bp
6. TBE Buffer (+ -) is 415bp,715bp
7. Agarose Gel, Tracking Dye (- -) is 415bp
8. DNA Stain "Gel Red" My results: (++)
9. Positive Control DNA
10. Base Pair Ladder
Procedure:
DNA Preparation:
1. Swirl 10 ml of saline in your mouth for 30 seconds.
2. Spit into cup and swirl to mix cheek cells.
3. Put 1000 ul of cheek cell solution into 1.5 ml microfuge tube.
4. Microfuge tube.
5. Pour off supernatant leaving 100 ul covering cell pellet.
6. Put 50 ul of cell suspension into tube of chelex.
7. Heat on heat block for 10 minutes.
8. Release pressure out of tube and microfuge.
9. Take 50 ul of supernatant from tube and place in new microfuge tube.
PCR:
1. Pipet 20 ul of master mix, 20 ul of primer mix, and 10 ul of DNA into new PCR tube.
2. Place in thermal cycler.
Gel:
1. 50 ml of 1x TAE an 1 g agarose
2. Heat solution until dissolved.
3. Pour in mold when cool.
4. 20 ml of DNA into new tube with 4 ml loading dye
5. Centrifuge and load 20 ul into well.
6. Record lane and gel number
Result:
My personal result was (++), meaning that both of my parents carry the Alu gene. After my results, I wonder if my 2 siblings also carry the Alu gene.