Genotyping using High Resolution Melting and Small Amplicons
Small amplicon genotyping uses primers that are
placed as close to the base of interest and develop amplicon’s of between 50
and 120bp. Depending on the Tm differences between the different genotypes this
method can give clear cut results. For those assays where the Tm differences
are very close, it is possible to use Idaho’s High Sensitivity
Mastermix. This mastermix contains internal calibrators to help clarify the
calls. The difference before and after using internal calibrators can be quite
dramatic – see image 1 & 2.
Without Calibrators
With Calibrators
Additional Information:
1) Small Amplicon Genotyping - Application Note
2) High Sensitivity Mastermix - Flyer
3) Improved Homozygous Detection - Poster
Images courtesy of Claire Taylor, Section of Epidemiology and Biostatistics, Leeds Institute of Molecular Medicine, Leeds.