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Instructions: You will need the following reagents and supplies: Tris/acetate/EDTA buffer pH 8.0 (TAE buffer) • Tris 40 mM Acetic acid 20 mM • EDTA 1 mM Agarose gel To make a 1% gel dissolve 1 gram agarose in 100 mL TAE buffer Add 10 µL of SYBR safe dye for imaging DNA Run in chamber with TAE buffer at 120 V for 25 minutes 200 µL thin-walled PCR tubes • 2X master mix containing buffer, polymerase, and all four nucleotide triphosphates • Nuclease free water • DNA template • Forward and reverse primers A thermocycler Procedure • Add the following PCR reagents added to a thin-walled PCR tube - 25 µL of 2X master mix - 2 µL primers (1 µM each) - 1 µL template - 22 µL water • You will prep both a control and your extracted DNA sample. • Place samples in a thermocycler using the following program 1. 95°C for 2 min 2. 95°C for 0.5 min 3. 56°C for 0.5min 4. 72°C for 1 min/1 kB of template 5. Repeat steps 2-4 40 times 6. 72°C for 10 min - final extension Pre-lab questions Name

          Instructions:
You will need the following reagents and supplies:
Tris/acetate/EDTA buffer pH 8.0 (TAE buffer)
• Tris 40 mM
Acetic acid 20 mM
• EDTA 1 mM
Agarose gel
To make a 1% gel dissolve 1 gram agarose in 100
mL TAE buffer
Add 10 µL of SYBR safe dye for imaging DNA
Run in chamber with TAE buffer at 120 V for 25
minutes
200 µL thin-walled PCR tubes
• 2X master mix containing buffer, polymerase, and all
four nucleotide triphosphates
• Nuclease free water
• DNA template
• Forward and reverse primers
A thermocycler
Procedure
• Add the following PCR reagents added to a thin-walled
PCR tube
- 25 µL of 2X master mix
- 2 µL primers (1 µM each)
- 1 µL template
- 22 µL water
• You will prep both a control and your extracted DNA
sample.
• Place samples in a thermocycler using the following
program
1. 95°C for 2 min
2. 95°C for 0.5 min
3. 56°C for 0.5min
4. 72°C for 1 min/1 kB of template
5. Repeat steps 2-4 40 times
6. 72°C for 10 min - final extension
Pre-lab questions
Name

Instructions:
You will need the following reagents and supplies:
Tris/acetate/EDTA buffer pH 8.0 (TAE buffer)
• Tris 40 mM
Acetic acid 20 mM
• EDTA 1 mM
Agarose gel
To make a 1% gel dissolve 1 gram agarose in 100
mL TAE buffer
Add 10 µL of SYBR safe dye for imaging DNA
Run in chamber with TAE buffer at 120 V for 25
minutes
200 µL thin-walled PCR tubes
• 2X master mix containing buffer, polymerase, and all
four nucleotide triphosphates
• Nuclease free water
• DNA template
• Forward and reverse primers
A thermocycler
Procedure
• Add the following PCR reagents added to a thin-walled
PCR tube
- 25 µL of 2X master mix
- 2 µL primers (1 µM each)
- 1 µL template
- 22 µL water
• You will prep both a control and your extracted DNA
sample.
• Place samples in a thermocycler using the following
program
1. 95°C for 2 min
2. 95°C for 0.5 min
3. 56°C for 0.5min
4. 72°C for 1 min/1 kB of template
5. Repeat steps 2-4 40 times
6. 72°C for 10 min - final extension
Pre-lab questions
Name

Added by Dale H.

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