Can Ampligase DNA Ligase be used in DNA polymerase buffers?
Ampligase DNA Ligase has very flexible buffer requirements, and will work in most DNA polymerase buffers. However, NAD must be added as a cofactor and is included in the Ampligase 10X Reaction Buffer. Q: To detect a point mutation using ligation amplification, how close must the point mutation be to the ligation site? A: We recommend that the point mutation be positioned at the ligation junction. The enzyme will detect a mutation one base away from the junction. However, at two bases away from the ligation junction, the discrimination between matches and mismatches drops dramatically. At three or more bases away, the mutation has little effect on ligation. Q: What is Repeat Expansion Detection (RED) and how is Ampligase DNA Ligase used in this procedure? A: RED is a ligation-based method of genetic screening that detects DNA regions composed of multiple trinucleotide repeats.6-8 Ampligase DNA Ligase is used to generate multimers of trinucleotide repeat oligonucleotide probes when cycle
