Mosaic DNA chip fabrication and its time-resolved fluorescence detection

2004 ◽  
Author(s):  
Quanguo He ◽  
Hong Chen ◽  
Jianxin Tang ◽  
Pengfeng Xiao ◽  
Nongyue He
Keyword(s):  
Fluorescence Detection ◽  
Dna Chip ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Chip Fabrication

Time-Resolved Fluorescence Detection of Mosaic DNA Chip

2006 ◽  
Vol 6 (1) ◽  
pp. 66-71 ◽  
Author(s):  
Quanguo He ◽  
Hong Chen ◽  
Libo Nie ◽  
Jianxin Tang ◽  
Pengfeng Xiao
Keyword(s):  
Fluorescence Detection ◽  
Perfect Match ◽  
Dna Chip ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Glass Slides ◽  
Base Mismatch ◽  
Detection Mode ◽  
Potential Alternative ◽  
Labeling Method

We demonstrated a time-resolved fluorescence (TRF) label and detection of mosaic DNA chip in this paper. We synthesized oligonucleotide sequences in situ on glass slides directly, and then sliced them up into small pieces and patched up the pieces bearing different sequences to generate a mosaic DNA chip. With multiple 4, 7-bis(chlorosulfophenyl)-1, 10-phenanthroline-2, 9-dicarboxylic acid (BCPDA, abbreviated as BCPDA) labeling method based on avidin-biotin amplification, we established a TRF detection format on the mosaic DNA chip. The detection method allows discriminatory signals for perfect match, one-base mismatch, two-base mismatch, and three-base mismatch by TRF labeled DNA hybridization, whereby Europium (III, Eu3+) was captured and released on the principle of complexation and dissociation interaction between BCPDA and Eu3+ solution when the BCPDA-tagged avidin and biotin-capped oligonucleotide sequence linked. The fluorescence spectra and related lifetimes were determined. We also compared the TRF detection mode with the conventional fluorescence one. These results showed the former is a potential alternative replacement of the latter, especially for labeling the mosaic DNA chip. The discovery is of fundamental interest and has significant implications to biochips and biosensors based on time-resolved-fluorescence detection.

Detection of Escherichia coli rRNA using target amplification and time-resolved fluorescence detection

1991 ◽  
Vol 5 (6) ◽  
pp. 467-472 ◽  
Author(s):  
Charlene E. Bush ◽  
Kurt M. Vanden Brink ◽  
David G. Sherman ◽  
W. Richard Peterson ◽  
Laura A. Beninsig ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fluorescence Detection ◽  
Time Resolved Fluorescence ◽  
Time Resolved

An automated PCR platform with homogeneous time-resolved fluorescence detection and dry chemistry assay kits

2008 ◽  
Vol 374 (2) ◽  
pp. 411-416 ◽  
Author(s):  
Virve Hagren ◽  
Piia von Lode ◽  
Anniina Syrjälä ◽  
Tero Soukka ◽  
Timo Lövgren ◽  
...  
Keyword(s):  
Fluorescence Detection ◽  
Time Resolved Fluorescence ◽  
Time Resolved

Time-resolved fluorescence detection of enzyme-amplified lanthanide luminescence for nucleic acid hybridization assays

1991 ◽  
Vol 37 (9) ◽  
pp. 1506-1512 ◽  
Author(s):  
E F Templeton ◽  
H E Wong ◽  
R A Evangelista ◽  
T Granger ◽  
A Pollak
Keyword(s):  
Alkaline Phosphatase ◽  
Nucleic Acid ◽  
Fluorescence Detection ◽  
Dna Hybridization ◽  
Nucleic Acid Hybridization ◽  
Time Resolved Fluorescence ◽  
Dot Blot ◽  
Time Resolved ◽  
Lanthanide Luminescence ◽  
Hybridization Assays

Abstract A new nonisotopic detection method based on time-resolved fluorescence for nucleic acid hybridization assays with alkaline phosphatase labels has been developed: enzyme-amplified lanthanide luminescence (EALL). EALL combines the amplification of an enzyme label with the sensitivity and background elimination of time-resolved fluorescence detection of lanthanide ion luminescence. The detection system for alkaline phosphatase makes use of a phosphorylated salicylic acid derivative that, upon dephosphorylation, gives a product capable of forming a luminescent terbium chelate. We demonstrate DNA hybridization assays by using two substrates, one for membrane and one for solution-based formats. Using the substrate that produces a more adhesive product allows performance of dot-blot and Southern blot assays on nylon membranes; results can be recorded with a time-resolved photographic camera system, or with an ultraviolet transilluminator-based system. Less than 4 pg of target sequence can be detected in a dot-blot assay after incubation with substrate for 2-4 h. DNA microwell-plate hybridization assays with the more soluble substrate/product pair can be quantified with time-resolved fluorescence plate readers, giving a similar detection sensitivity. EALL is thus a practical time-resolved fluorescence-based alternative to other detection systems for DNA hybridization assays.

scholarly journals Gene expression analysis with an integrated CMOS microarray by time-resolved fluorescence detection

2011 ◽  
Vol 26 (5) ◽  
pp. 2660-2665 ◽  
Author(s):  
Ta-chien D. Huang ◽  
Sunirmal Paul ◽  
Ping Gong ◽  
Rastislav Levicky ◽  
John Kymissis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Fluorescence Detection ◽  
Gene Expression Analysis ◽  
Time Resolved Fluorescence ◽  
Time Resolved
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