DNA Methodologies. Sterilization Clean the workstation with alcohol and bleach. Autoclaving and ultraviolet light (UV radiation). Consumables and reagents. Equipment Pipettes- P20, P200, P1000 Block heater Vortex Centrifuge PCR machine Electrophoresis box and power supply
Genetic Analyzer- mtDNA sequencing and STR analysis
Analysis of Results
1. Epithelial cells are lysed with a mild extraction buffer containing Sodium Dodecyl Sulfate (SDS).
2. Centrifugation to pellet sperm cells, DNA from epithelial cells is in the supernatant.
3. Lyse sperm cells using Dithiothreitol (DTT).
During the exponential expansion of the PCR the amount of product produced is proportional to the amount of template. Here we show the total amount of product following 32 cycles.
Develop a standard curve