Changes

#Spin down plate 1 min at 3000 RPM in an empty 96 well plate using the Lin lab centrifuge#Press power button.#Open tray and put plate into tray.#Click on “Shortcut to Sybr plate” “7300 System Software” icon.#Click on “Instrument” tab and change volume of rxn. to 20uLopen then in the directory Templates select SYBR Plate or SYBR Plate (no dissociation). With new primers, may want to run the dissociation step to see if one peak (or more) is generated by the primers. This step adds about an extra hour to the run time. Can eliminate this step for primers which have already been analyzed in this way.#Click on “Instrument” tab and check that reaction volume is 20uL.#Go to File menu and then Save As. Go up one folder to ABI folder then to the Results folder, then save in the Saltiel folder (in my folder).#Close with the program (Shortcut to Sybr plate window).#Reopen date as the program through the 7300 System Software icon. Open the new saved file and the 7300 system should then initializefilename.#Under the Instrument tab, hit “Start.” Note: #Wait until a estimated run time is shown. It takes a few minutes for the run to begin after “Start” is hit. Sometimes the run time is incorrectly calculated by the software.