03 - Chick Check: The Genetics Behind Blue Chicken Eggs
Module I: Amplification of DNA
GENTLY MIX
TC
• 20 μL Primer Mix • 5 μL DNA Sample • PCR EdvoBead™
SPIN
Before you begin, OBTAIN the following materials: labeled DNA tem- plates, PCR EdvoBeads™, PCR Master Mix, and PCR tubes.
LABEL PCR tubes.
1. 2. 3.
PLACE all materials on ice.
ADD to each tube in the following order: • 20 µL LyphoPrimer • 5 µL DNA template (use water for negative control) • 1 PCR EdvoBead 4. MIX each tube by gently flicking or brief vortexing. 5. CENTRIFUGE briefly to collect the sample at the bottom of the tubes. 6. PLACE tubes and the thermal cycler and RUN the following program: • Initial denaturation: 95°C for 5 minutes • 95° C for 30 seconds • 59° C for 30 seconds • 72° C for 30 seconds • Final extension: 72°C for 10 minutes. 7. After PCR, HOLD tubes at 4°C. PROCEED to Module II: Agarose Gel Electrophoresis. OPTIONAL STOPPING POINT: The PCR samples may be stored at -20° C for electrophoresis at a later time. 35 cycles }
RELATED VIDEO: Ingredients for PCR Success
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