Topics in Fluorescence Spectroscopy: DNA Technology by Steven A. Soper, Clyde Owens, Suzanne Lassiter (auth.),
By Steven A. Soper, Clyde Owens, Suzanne Lassiter (auth.), Joseph R. Lakowicz (eds.)
During the previous 15 years, there was amazing development within the research and manipulation of DNA and its use in nanotechnology. DNA research is ubiquitous in molecular biology, scientific diagnostics, and forensics. a lot of the readout know-how is predicated on fluorescence detection. This quantity comprises contributions from many specialists within the box who current an outline of many features of DNA know-how. those chapters supply an knowing of the underlying rules and know-how, instead of an exhaustive evaluate of the literature. Written in a transparent straight forward sort, this booklet is a superb creation for any scientist to using fluorescence in DNA analysis.
DNA Technology is a necessary analyzing for all teachers, bench scientists, and execs wishing to exploit the most recent and maximum during this consistently rising field.
*Comprehensive evaluate of the complexities of DNA research,
*Covers themes of common curiosity to a large box of scientists,
*Accessible application in featuring cutting-edge DNA know-how,
*Chapters authored by means of key figures within the box.
Read or Download Topics in Fluorescence Spectroscopy: DNA Technology PDF
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Additional info for Topics in Fluorescence Spectroscopy: DNA Technology
As such, the red-dyes are used at higher concentrations during DNA polymerization to circumvent poor excitation. 13–18 While the focus of a subsequent chapter in this book will deal with this topic, it is informative to briefly introduce these energy transfer (ET) primers at this time in the context of DNA sequencing. 14. The donor dye in this case was FAM, which could be excited with the 488 nm line of an Ar ion laser. The acceptor dyes were either FAM, JOE, TAMRA, or ROX. The donor (FAM) was attached to the sequencing primer on the 5' end during the solid-phase DNA synthetic preparation of the M13 (-40) sequencing primer using phosphoramidite chemistry.
Soper et al. centage of crosslinker, N,N'-methylenebisacrylamide in the presence of a catalyst accelerator-chain initiator mixture. The porosity of the gel is determined by the relative proportion of acrylamide monomer to crosslinking agent. For reproducible fractionation of DNAs, they must be maintained in a single-strand conformation, which is accomplished by adding denaturants, such as urea or formamide, to the sieving gel. , electrophoretic mobility) observed in the sequencing process. Any DNA fragments to be fractionated will inevitably encounter the gel network of polymer threads.
These components are solid-state allowing the detector to be run for extended periods of time requiring little maintenance or operator expertise. Near-IR fluorescence can be a very attractive detection strategy in gel sequencing because of the highly scattering medium that the separation must be performed in. Due to the intrinsically lower backgrounds that are expected in the near-IR compared to the visible, on-column detection can be performed without DNA Sequencing Using Fluorescence Detection 33 sacrificing detection sensitivity.