qPCR - Halley cDNA Check

Ran qPCR on Halley’s cDNA to see if I could get them to work. She has been getting high levels of fluorescence at the initiation of the qPCR cycling that shouldn’t be there. Master mix calcs and plate layout can be seen here. http://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20121128%20qPCR%20Layout.jpg

Cycling params can be found in the qPCR Data File (see Results).


qPCR Data File (Opticon2)


High levels of initial fluorescence are present in both sets of cDNA samples, while the NTC sample does not exhibit this behavior, suggesting the template is to blame. Have suggested to Halley to make new cDNA using the correct recipe, instead of the FISH441 recipe she had been using.