High Resolution Melting has a number of requirements if the user is to have any success with the method. You'll need good quality amplification, an instrument capable of high density data collection and exceptional thermal accuracy and precision. You will also need a dye that has the capacity to saturate your dsDNA molecule.
LCGreen Plus Dye:
Designed specifically for High Resolution Melting applications, LCGreen Plus has been shown through experimentation to provide consistent saturation of dsDNA to offer reliable and sensitive data about the melting properties of the dsDNA in your experiment. Figure 1 shows a theoretical idea of the differences between a saturation dye and a non saturation dye.
Figure 1: As the dsDNA dissociates so the dye (SYBR Green) re-attaches in another section of the molecule. This stops a decrease in flourescence. While with LCGreen Plus as the dsDNA dissociates so the flourescence decreases accordingly. This repeatable time after time and this makes High Resolution Melting a possible tool for mutation scanning, genotyping ........
Idaho provide LCGreen Plus Dye in two formats. One a ready to use 10x concentrated dye and also as part of a PCR Mastermix that comes at a 2.5x concentration and contains everything required for PCR except for dsDNA and primers.
Additional Information.
1)
LCGreen Plus Dye - Flyer
2)
LCGreen PCR Mastermix - Flyer
3)
LCGreen Plus High Sensitive PCR Mastermix - Flyer
4)
LCGreen Plus - Application Note
5)
Comparison of various Mastermixes and dyes for HRM - Poster