1, Why it is necessary to chelate the metal ions from a solution during the boiling lysis step at 100°C. What would happen if you do not use a chelating agent such as Insta gene matrix?
Answer: Addition of chelating agent helps to chelate metal ions (e.g. Mg2+) that are used as cofactor for DNase. By removing these metal ions, DNase is rendered inactive as a result the template DNA stays intact.
2. What is needed from the cells for PCR?
Answer: For PCR amplification to occur the genomic material (DNA) from the cell's nuclei is required to be used as a template for amplification.
3. What structures must be broken to release the DNA from the cell?
Answer: Cell wall and Cell membrane
4. Why is the DNA stored cold with the Insta gene matrix after boiling the samples?
Answer: Low temperature ensures that any background enzymatic activity that could degrade the DNA is reduced.