|
To realize the potential of HRM, an instrument system requires:
- The absolute minimum of well-to-well thermal and optical variation
- Exquisite control of sample temperature
- Uniformity of thermal transition (ramp) rates across all samples
- High intensity illumination
- High sensitivity optical detection
- A fast data acquisition rate
- Advanced software designed especially for HRM data analysis
By virtue of its unique rotary design, the Rotor-Gene 6000 instrument excels in each respect and is in a class of its own for HRM capability.
Corbett Life Science offers two instruments designed to support HRM; the Rotor-Gene 6000 2-plex+HRM and the Rotor-Gene 6000 5-plex+HRM.
Increasing interest in HRM has prompted some manufacturers to claim their instrumentation supports HRM. However, true HRM can reliably genotype homozygous Class 4 SNPs (A>T), a capability beyond the reach of conventional block-based designs. We recommend a side-by-side comparison with the Rotor-Gene before making a purchasing decision. Be sure to arrange replicate samples all across the block and rotor to compare actual precision.
Click for a publication contrasting HRM on the Rotor-Gene to the Roche LightCycler 480 instrument
Click to view a detailed comparison of HRM on the Rotor-Gene and the Idaho Technology LightScanner™
"Resolution varied greatly among instruments with a 15-fold difference in Tm SD (0.018 to 0.274°C) and a 19-fold (LC Green Plus) or 33-fold (SYBR Green I) difference in the signal-to-noise ration. These factors limit the ability of most instruments to accurately genotype single-nucleotide polymorphisms by amplicon melting. Plate instruments (96-well) showed the greatest variance with spatial differences across the plates."
Hermann et al 2006
"The ability of most instruments to accurately genotype single-base changes by amplicon melting was limited by spatial temperature verification across the plate."
Herrmann et al 2007(b) |
