Fluorescent sequencing

Yamakawa, H., and Ohara, O. (1997). A DNA cycle sequencing reaction that minimizes compressions on automated fluorescent sequencers. Nucleic Acid Res. 25(6), 1311-1312. 

A regiment of scientists and technicians al Caltech and Applied Biosystems, Inc., invented the automated DNA fluorescence sequencer. 

The composition and conditions of use for ten selective staining reagents, when used in conjunction with two-dimensional solvent systems (H9), permits the unambiguous identification of seventy-six compounds of biochemical interest. Twenty-seven of these compounds may be identified specifically by a single reagent which is capable of producing a unique color (or fluorescence) sequence when the spot is viewed under successive conditions. 

RFLP, fluorescent sequencing technology SSCP analysis, radiolabelling techniques... 

Chen, C.T. Wagner, H. Still, W.C. Fluorescent, sequence-selective peptide detection by synthetic small molecules. Science 1998, 279, 851-853. 

Scherer N F, Carlson R J, Matro A, Du M, Ruggiero A J, Romero-Rochin V, Cina J A, Fleming G R and Rice S A 1991 Fluorescence-detected wave packet interferometry time resolved molecular spectroscopy with sequences of femtosecond phase-locked pulses J. Chem. Rhys. 95 1487... 

Nucleic acid (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)) probes utilize labeled, ie, radioactive, enzymatic, or fluorescent, fragments of DNA or RNA (the probe) to detect complimentary DNA or RNA sequences in a sample. Because the probe is tailored for one specific nucleic acid, these assays are highly specific and very sensitive (45). 

In recent years, automated DNA sequencing machines capable of identifying about 10 bases per day have become commercially available. One clever innovation has been the use of fluorescent dyes of different colors to uniquely label the primer DNA introduced into the four sequencing reactions for example, red for the A reaction, blue for T, green for G, and yellow for C. Then, all four reaction mixtures can be combined and run together on one electrophoretic... 

This group was developed as part of a scheme to prepare fluorescent tags to be used in DNA sequencing. Deprotection is accomplished by irradiation at 360 nm to release the NVOC group, which then sets up the system to form a diketopiper-azine while releasing the alcohol. ... 

When reaction is complete, the product consists of a mixture of DNA fragments of all possible lengths, each terminated by one of the four dye-labeled dideoxyribonucleotides. This product mixture is then separated according to the size of the pieces by gel electrophoresis (Section 26-2), and the identity of the terminal dideoxyribonucleotide in each piece—and thus the sequence of the restriction fragment—is identified simply by noting the color with which the attached dye fluoresces. Figure 28.8 shows a typical result. 

A diagnostic method using fluorescence labeled DNA probes to detect and quantify the number complementary chromosomal sequences on a cellular resolution. A related technique that also allows assessment of gene amplifications, but without precise quantification of copy numbers is the chromogenic in situ hybridization (CISH). Here, instead of a fluorescent dye an enzyme that can generate a colored precipitate in the tissue samples is coupled to the DNA probe. 

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