Key Points on Using Step One Plus for qPCR

Powering On

• Press the blue power button on the bottom left to turn on the machine.
• It takes a moment to boot up.

Machine Overview

• Similar to Quant Studio systems but has an extra dye.
• A dye guide will be posted on the cork board, indicating which dyes are calibrated for each machine and their compatibility.

Setting Up Protocols

• Shortcuts for protocols can be created.
• Pre-set for standard and fast protocols; ensure to select the correct one.
• To modify a protocol:
  • Click on each stage to adjust settings.
  • Options include:
    • Ramp rates
    • Cycle durations
    • Data point collection timings
• Adding Melt Curve:
  • If not configured, you can add a stage for a melt curve.
  • Look for a "c" symbol, indicating a melt curve may already be present.
• Saving Protocols:
  • Hit save to overwrite the existing experiment.
  • New shortcuts will appear in the experiment menu.

Loading Plates

• Remove the lid to access the well plate area.
• Ensure the assay plate fits properly in the Applied Bio Systems machine.
• Use an insert for deeper well plates to provide extra height.

Running the Experiment

• After placing the assay plate, close the lid.
• Hit start run to begin the experiment.

Collecting Results

• At the end of the run, you can export results:
  • Insert a USB key into the machine.
  • Navigate to main menu > collect results to export data.

Powering Down

• After data collection, remove the assay plate and replace the lid.
• Turn the power off on the machine.