![]() “Skip” allows you to skip to next step (e.g. Choose to enter sample names now or click “Finish” and enter sample names later Run will commence, while the experiment is running you will see: Raw Channel - fluorescence collected cycle by cycle Temperature - profile as temperature cycles Profile Progress – tells you time remaining, shows you where you are. To save as a template open template folder C:\Program Files\Rotor-Gene 6000 Software\Templates and save as template file (*.ret) Run files are *.rex Gain can be compared to the aperture on a camera and is used to fit data on raw scale If your starting fluorescence signal is weak the gain should be high, if strong the gain should be low Many people just accept default gain and go to “Next” on this screen, others optimise the gain for each run Click on “Gain Optimisation” to beginĬlick on “Optimise Acquiring” and “Perform Calibration Before 1st Acquisition” If you have different colours in different positions click “Edit” The colour you want to use must be in the tube position for calibration to be successful Don’t set gain on no template controlsĬlick on “Start Run” Save file name in directory of your choice Default nomenclature has name of template file, date and version (1 for 1st run that day etc) With SYBR use “Melt” to look for primer-dimer or non-specific product Melt should start from acquiring temperature If you start the melt from annealing temp the melt may have a shoulder on it Use default settings, make sure you are acquiring data (blue dots) click on “Edit Profile” to change run parameters section to be altered is greyed out, click on “Hold Temperature” to change temperature, click on “Hold Time” to change time Always read manufacturer’s instructions regarding Hot start Taq activation timesĬlick on “Cycling” to adjust the cycling parameters Default cycle number is 40 To change a step click on the temperature and adjust then click on the time and adjustĭata is acquired each cycle (blue dots) No blue dots = no data = very bad Acquire at the end of the extension phase Click on “Aquiring to Cycling A” to select colours Move the colours you want from “Available Channels” to “Acquiring Channels” using arrows Use Green channel for SYBR green Insert user name, any notes (primer conc, primer sequence mix used etc) and volume required (10-25uL) This info is saved and can be printed in reports Click “Next” Select the rotor you are using, put the rotor into the machine with the locking ring attached The locking ring tick box must be ticked to proceed Click “Next” ![]() ![]() Getting Started…… Click on Rotor-Gene icon Click on advanced tab for step by step run set up Choose run template depending on chemistry Dual labeled probes = hydrolysis probes SYBR = intercalating dyes The world’s only real-time rotary thermo-optical analyser Corbett LIFE SCIENCE Getting Started on the Rotor-Gene-6000 Jennifer McMahon, PhD
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