Ran PCR using fresh working stock of primers.
Adjusted primer volumes being used. Recipe from Lisa (from the class) were only using 0.1uL of each primer per reaction.
Also, tried additional template DNA to determine if the template I’ve been using has become degraded. Additional template were two other WS plasmid curves from 20120731; 3e7. Master mix calcs are here.
All samples were run in duplicate.
Success!!!!! Unfortunately, I can’t say definitely if the previously used working stocks were bad or if the primer amount being used was too low (I suspect the latter), but it works and that’s all that matters. Will proceed with finding the limit of detection for this assay.