Document Detail


A highly uniform UV transillumination imaging system for quantitative analysis of nucleic acids and proteins.
MedLine Citation:
PMID:  18398876     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The digital fluorescent imaging for documentation and analysis of gel electrophoretic separations of nucleic acids and proteins is widely used in quantitative biology. Most fluorescent stains used in postelectrophoretic analysis of proteins and nucleic acids have significant excitation peaks with UV light (300-365 nm), making midrange UV (UV-B) as the excitation source of choice. However, coupling quantitative CCD imaging with UV is difficult due to lack of uniformity found in typical UV transilluminators. The apparent amount of those macromolecules depends on the position of the gel band on the imaging surface of the transilluminator. Here, we report the development and validation of a highly uniform UV transillumination system. Using a novel high density lighting system containing a single lamp formed into a high density grid, an electronic ballast, a phosphor coating, and a bandpass filter to convert 254 nm light produced to 300-340 nm, uniformity of </=5% CV has been achieved, compared to >80% CV observed in typical UV transilluminators. This system has been used for the quantitative analysis of electrophoretically separated nucleic acids and proteins (CV</=8%) without uniformity correction by software, and continues to outperform typical transilluminators when a uniformity correction is applied.
Authors:
Bulbul Chakravarti; Melissa Louie; Wongrat Ratanaprayul; Alpan Raval; Sean Gallagher; Deb N Chakravarti
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Proteomics     Volume:  8     ISSN:  1615-9861     ISO Abbreviation:  Proteomics     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-05-05     Completed Date:  2008-09-15     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101092707     Medline TA:  Proteomics     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1789-97     Citation Subset:  IM    
Affiliation:
Keck Graduate Institute of Applied Life Sciences, CA, USA. bulbul_chakravarti@kgi.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
DNA / analysis
Electrophoresis, Polyacrylamide Gel
Equipment Design
Fluorescent Dyes / pharmacology
Nucleic Acids / analysis*
Proteins / analysis*
Proteomics / instrumentation*,  methods*
Reproducibility of Results
Serum Albumin, Bovine / chemistry
Spectrophotometry, Ultraviolet / methods
Staining and Labeling
Transillumination
Ultraviolet Rays
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 0/Nucleic Acids; 0/Proteins; 0/Serum Albumin, Bovine; 9007-49-2/DNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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