Synthesis and Application of 3′-Amino-Dye-Terminators For Dna Sequencing

The use of dideoxynucleotide triphosphates labeled with different fluorescent dyes (dye terminators) is the most versatile method for automated DNA sequencing. However, variation in peak heights reduces base-calling accuracy and limits heterozygous allele detection, favoring use of dye-labeled primers for this purpose. We have discovered that the addition of a manganese salt to the PE Applied Biosystems dye-terminator sequencing kits overcomes these limitations for the older rhodamine dyes as well as the more recent dichloro-rhodamine dyes (dRhodamine and BigDyes). Addition of manganese to reactions containing dRhodamine-based dye terminators produced the highest base-calling accuracy. This combination resulted in the most uniform electropherogram profiles, superior to those produced by BigDye terminators and published for dye primers, and facilitated detection of heterozygous alleles.

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Two-Step Cycle Sequencing Improves Base Ambiguities and Signal Dropouts in DNA Sequencing Reactions Using Energy-Transfer-Based Fluorescent Dye Terminators

Molecular Biotechnology ◽

10.1385/mb:17:2:135 ◽

2001 ◽

Vol 17 (2) ◽

pp. 135-142 ◽

Cited By ~ 7

Author(s):

Long Wen

Keyword(s):

Energy Transfer ◽

Dna Sequencing ◽

Fluorescent Dye ◽

Step Cycle ◽

Cycle Sequencing ◽

Base Ambiguities ◽

Dye Terminators

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Imaging polymers, proteins and dna in aqueous solutions with the atomic force microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100152136 ◽

1989 ◽

Vol 47 ◽

pp. 32-33

Author(s):

S.A.C. Gould ◽

B. Drake ◽

C.B. Prater ◽

A.L. Weisenhorn ◽

S.M. Lindsay ◽

...

Keyword(s):

Amino Acids ◽

Dna Sequencing ◽

Aqueous Solutions ◽

Atomic Force Microscope ◽

Piezoelectric Crystal ◽

Atomic Force ◽

Structure Of Molecules ◽

Optical Lever

The atomic force microscope (AFM) is an instrument that can be used to image many samples of interest in biology and medicine. Images of polymerized amino acids, polyalanine and polyphenylalanine demonstrate the potential of the AFM for revealing the structure of molecules. Images of the protein fibrinogen which agree with TEM images demonstrate that the AFM can provide topographical data on larger molecules. Finally, images of DNA suggest the AFM may soon provide an easier and faster technique for DNA sequencing.The AFM consists of a microfabricated SiO2 triangular shaped cantilever with a diamond tip affixed at the elbow to act as a probe. The sample is mounted on a electronically driven piezoelectric crystal. It is then placed in contact with the tip and scanned. The topography of the surface causes minute deflections in the 100 μm long cantilever which are detected using an optical lever.

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