University:
Duke University
Course:
BIOLOGY20 | General Biology
Academic year:

2024
Views:

332

Pages:

1
Author:

Rebecca B.

PCR and ELECTROPHORESIS Material : - Agarose - TBE/ TAE 0.5x - DNA sample - Markers - Loading dye 6x - PCR kits - Aquabidest - Primary - Ethidium bromide Tool: - PCR Machine (Thermocycler) - Micropipettes (1-10 uL, 2-20 uL, 20-100uL, 100-1000uL) - Tips (white, yellow, blue) - microcentrifuge tubes measuring 0.2 mL and 1.5 mL - Electrophorator - UV transilluminator - Handscoon - Camera Ways of working: 1. Prepare materials for PCR mastermix (sample DNA, PCR kit, primers and aquabidest). 2. Put the PCR mastermix ingredients into a 0.2 ml tube (2 µl DNA, 10 µl PCR kit, 1 µl primer, and aquabidest until the final volume of the mixture reaches 20 µl). 3. Close the tube tightly and insert it into the PCR machine. 4. Set the temperature, time and cycle on the PCR machine. 5. Start the PCR machine. 6. PCR products are analyzed using agarose gel electrophoresis. 7. Read the gel electrophoresis results of the PCR product. 8. Report the results you got in the practicum.