A modified restriction display PCR method in sample-labelling of DNA microarray.

Ling Li; Wen Li Ma; Ji Zhu; Rong Shi; Cui Hua Liu; Jin Kun Chen; Wen Ling Zheng

doi:10.1016/j.jviromet.2003.09.013

Home/Archive/PubMed

PubMed · 14599681

A modified restriction display PCR method in sample-labelling of DNA microarray.

Ling Li·Wen Li Ma·Ji Zhu·Rong Shi·Cui Hua Liu·Jin Kun Chen·Wen Ling Zheng

View original source ↗Export metadata ↓

Abstract

The restriction display PCR is a useful technique for studying the diversity of gene expression. This method involves ligating the digested genes with adapters and amplifying the gene fragments by PCR using universal and selective primers. In this study, we improved this restriction display PCR method by using Cy3-UP, a fluorescently labelled universal primer, in place of Cy3-dCTP in sample-labelling for DNA microarray. The results show that this new method increases significantly the sensitivity of the assay, and will have a wide application in the DNA microarray field.

Explore related subjects

Carbocyanines DNA Primers DNA, Viral Fluorescent Dyes HIV-1 Humans Oligonucleotide Array Sequence Analysis Polymerase Chain Reaction Specimen Handling Evaluation Study Journal Article Research Support, Non-U.S. Gov't

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Ling Li, Wen Li Ma, Ji Zhu, Rong Shi, Cui Hua Liu, Jin Kun Chen, Wen Ling Zheng. 2003. A modified restriction display PCR method in sample-labelling of DNA microarray.. https://doi.org/10.1016/j.jviromet.2003.09.013

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

Chiral transformation of achiral J-aggregates of a cyanine dye templated by human serum albumin.

Explore the source record for details and available documents.

Carbocyanines↗

PubMed2006

Real-time quantification of RNA polymerase activity using a "broken beacon".

A novel assay using a hybridization-based method was developed for real-time monitoring of RNA synthesis. In this work, a "broken beacon" in which the fluor and quencher were located on separate but complementary oligonucleotides was used to quantify the amount of RNA production by T7 polymerase. The relative lengths of the fluor-oligo and quencher-oligo, and their relative concentrations were optimized. The experimentally determined limit-of-detection was approximately 45 nM. The new assay was compared to the "gold-standard" radiolabel ([(32)P]NTP incorporation) assay for RNA quantification. While the broken beacon assay exhibited a higher limit of detection, it provided an accurate measure of RNA production rates. However, the broken beacon assay provided the significant analytical advantages of (i) a real-time and continuous measurement, (ii) no requirement for the use of radiolabels or gel-based analysis, and (iii) substantial time and labor savings.

Carbocyanines↗

PubMed2006

Characterization of DNA hybridization on partially aminated diamond by aromatic compounds.

Here, we report a novel method of micropatterning oligonucleotides via aromatic groups as linkers on partially amino-terminated diamond and the inherence on subsequent hybridization. The covalent immobilization of probe oligonucleotides and characterization of immobilized probe oligonucleotides with carboxylic compounds were investigated by X-ray photoelectron spectroscopy (XPS). To confirm the effects of linker flexibility in a low amino group on diamond for probe oligonucleotides, three kinds of dicarboxylic compound--adipic acid, terephthalic acid, and trimesic acid--were used for immobilization of probe oligonucleotides, like linkers; and these oligonucleotides were hybridized with target oligonucleotides labeled with Cy 5 on the micropatterned diamond surface. The hybridization intensities determined by epifluorescence microscopy were compared and analyzed.

Carbocyanines↗