What we do

 Top Software

High-Resolution Melting
See updated instrument comparison data

Why do we need a special machine ?

The accuracy of mutation detection and genotyping depends on the quality of melting curves generated by the instrument. Unfortunately, most real-time PCR machines are inadequate in this respect. We developed two instruments for high-resolution melting, the HR-1 and the LightScanner. To date, HR-1 has the highest resolution of all commercial instruments. It uses glass capillary tubes for optimal temperature control. After PCR, melting analysis is complete in 1-2 min. The LightScanner is the best solution for high-throughput analysis on 96 or 384-well plates. Paired with standard plate thermal cyclers, the throughput of the system reaches over 4,000 samples an hour on the 384-well LightScanner if enough thermal cyclers are available (melting of one plate is achieved in 5 min.)
data density click image to enlarge

data precision click image to enlarge

How much resolution is "high resolution"?

Based on functional testing of instruments currently available, acquisition of 10 datapoints per degree C during melting operations is probably the minimum requirement to allow some level of mutation scanning. However, temperature precision is equally important, and higher data density is probably required for detection of sequence variants that exist at low allelic fractions.

The conventional real-time PCR instrument (LightCycler 2.0) only acquires one datapoint per degree C when the carrousel is fully loaded (32 samples), while the HR-1 acquires 200 datapoints/ °C (figure right).

Noise or scanning precision (measured as the standard deviation in Tm after temperature shifting) was obtained for two other commercial instruments. The HR-1 instrument has the highest data density and the highest precision. Of the high-throughput systems, the LightScanner has the highest precision (bottom right). Further details can be obtained from Herrmann et al 2007. For updated data, see top of this page.

Instruments for High-Resolution Melting Anaysis line up


© 2006- Wittwer Lab. All Rights Reserved.