Used COX primers (SR IDs 1060, 1061) and cDNA from 20080327, which consisted of 7 control gigas gill and 7 vibrio-exposed (24hrs) gigas gill samples, labeled as C# and VE#, respectively. The experiment was a 24hr. exposure live Vibrio vulnificus, parahaemolyticus Cf = 2.055x10^11 (6.85x10^7 Vibrio cells/oyster). Note: Used a free sample of 2x Brilliant III Ultra Fast SYBR Green QPCR Master Mix (Stratagene) for this qPCR. Mixed components and set up cycling params according to the manufacturer’s recommendation for the BioRad CFX96.
Master mix calcs are here. Plate layout, cycling params, etc. can be see in the qPCR Report (see Results).
Results:
[ qPCR Report (PDF)(https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20101213%20qPCR%20Report-01.pdf).
PCR Miner analysis is here. There appears to be an increase in COX expression in samples exposed to Vibrio sp. (see graph below), however, I have not determined if the results are significant.
