Snapshot kits

FIGURE 6.4 Principle of the Snapshot Kit. (a) Primers specific to a sequence of PCR product up to a position in front of the single-nucleotide polymorphism binds to PCR product. In the reaction, all four fluorescence dye-labeled ddNTPs are present, (b) Enzymatic elongation stops directly after incorporation of ddNTP. (c) After cleanup of the reaction, the elongated dye-labeled primer is analyzed by capillary electrophoresis. 

Applied Biosystems, ABI Prism SNapShot Multiplex kit, http //www.applied-biosystems.com. 

Thaw the reagents in ABI PRISM SNaPshot Multiplex Kit on ice just before the reaction and keep the reaction mix in dark and on ice. 

SNaPshot multiplex ready reaction mix contained in the ABI Prism SNaPshot multiplex kit (Applied Biosystems, Foster City, CA). 

Several modes of detecting a single base extension have been investigated, including measuring the incorporation of fluorescent, haptenated, or radioactive ddNTPs [6, 7] or gel electrophoresis based-detection of fluorescent primers extended by non-fluorescent nucleotides [8]. Recently, the Applied Biosystems SNapshot lYimer Extension Kit was introduced. In this assay, a primer is extended by one or more fluorescent- labelled dideoxynucleotides with subsequent detection in a fluorescent-based DNA sequencer. Several primers can be analysed within one lane of the DNA sequencer. 

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