BLAT: Molecular and Immunological Methods. Lyle McMillen Contact: firstname.lastname@example.org. Molecular and Immunological Methods. 2 basic approaches will be covered, both based on specific interactions found in vivo Nucleic acid specificity (DNA and RNA binding)
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Taq polymerase – 94 kDa monomer
Dichlororhodamine dyes are used to label ddNTP’s in a dideoxy terminator reaction.
Each ddNTP is labelled with a particular variant dye, with different emission wavelengths (i.e. Different colour), resulting in a single reaction generating random fragments, with each fragment labelled with a dye that corresponds to the terminal base.
This trace is easily interpreted, with each peak corresponding to the terminal base on the labelled DNA fragment.
A number of high throughput sequencing technologies are being developed, with the goal of sequencing millions of bases very rapidly.
NB: This plot is normalised to the mutant allele melting curve.
A 40000 spot two-colour oligo microarray.