Question

Now that the student has the dNTP stock ready, they are ready to proceed with their PCR. Since this is the first time the student is amplifying this particular gene from this new organism, what is a good control to use to ensure that their DNA isolation worked?

Answer 1

• The use of formalin fixative stems from its ability to cross-link proteins, however it has also been reported to induce methylol modifications and acid-mediated hydrolysis which can confound genotyping analysis
• DNA extraction is usually performed using commercially available kits. Variation between manufacturers and even lot to lot from the same manufacturer is common
• DNA quantitation is not performed by all laboratories but can be useful to ensure that DNA isolates meet minimum concentration for use in an assay. Measurement systems can be confounded by low sample yield and sample fragmentation.
• Pre-analytical processes are not always automated and great skill is required to successfully perform a DNA extraction from an FFPE sample without losing material.