Dye-labeled primers

Myburg, A.A., D.L. Reminglon, D.M. O Malley, R.R. Sederoff, and R.W.WhetLen, High-throughput AFLP analysis using infrared dye-labeled primers and an automated DNA sequencer. Biolechniques, 2001. 30(2) 348-52, 354,356-7. 

The dideoxy DNA sequencing method begins with the denaturation of double-stranded DNA (dsDNA) into single-stranded DNA. The ssDNA is then annealed with a fluorescent dye-labeled primer, which is an oligodeoxynucleotide 20 bases long. The heteroduplex formed is then incubated in four separate reactions. 

Design and synthesis of fluorescence energy transfer dye-labeled primers and their application for DNA sequencing and analysis Anal. Biochem. 231 131 0... 

FIGURE 1.5 Dye-labeled DNA fragments are produced by PCR with dye-labeled primers (a). The size of two different alleles (b) with 7 or 5 repeats (4 bp) differs from that of 8 bp. By coseparation of a size standard (c, black Unes) and an allelic ladder (c, gray Unes), the size can be determined and the alleles determined correctly. 

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. 

The use of donor-acceptor pairs allows for the equalization of emission intensity at a common excitation wavelength. The use of energy transfer fluorescent dye labeled primers offers improved sensitivity with respect to single dye labeled primers [90, 101, 109, 110]. Examples of energy transfer schemes are shovm in Fig. 14.18. 

Ju, J. Y., Ruan, C. C., Fuller, C. W., Glazer, A. N., and Mathies, R. A. Fluorescence energy-transfer dye-labeled primers for DNA-sequencing and analysis. Proceedings of the National Academy of Sciences of the United States of America 92, 4347-4351 (1995). 

Ansorge, W Voss, H. Stegemann, J. Wiemaim, S. Multiplex DNA sequencing using dye-labeled primers. Ger. Offen. DE 19515552,1996. 

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