Literature DB >> 15013421

Dextran sulfate provides a quantitative and quick microarray hybridization reaction.

Wei-Chi Ku1, Wai Kwan Lau, Yu-Tien Tseng, Chi-Meng Tzeng, Sung-Kay Chiu.   

Abstract

Microarray technology is a powerful tool to speed up genomics study, yet many technical aspects need to be improved. The hybridization reaction of microarray experiments is carried out for 16h or overnight in order to obtain reasonably strong signals for analysis in the presence of high salt buffer, like SSC. However, the quantitative aspect of microarray hybridization has seldom been investigated. In this study, we showed that higher overall signals from hybridization were achieved in a buffer system containing dextran sulfate, which can accelerate the kinetics of reaction by increasing the local concentration of the reactants. The dextran sulfate containing hybridization solution increases the reaction 4-fold (median) for cDNA microarray and 29-fold for oligonucleotide microarray. More importantly, the solution also provides a quantitative hybridization reaction, where the hybridization signals are proportional to the abundance of transcript added. The enhancement in the kinetics of hybridization is due to both dextran sulfate and formamide present in the solution, but the effect is not due to the higher temperature used during the reaction. With a slightly longer reaction time the hybridization reaction with the solution allows the detection of hybridization signals from rare transcripts that is not possible with regular hybridization buffers. With appropriate washing, the enhancement of kinetics by the solution does not increase the background signals at all, allowing higher signal-to-noise ratios to be achieved.

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Year:  2004        PMID: 15013421     DOI: 10.1016/j.bbrc.2004.01.013

Source DB:  PubMed          Journal:  Biochem Biophys Res Commun        ISSN: 0006-291X            Impact factor:   3.575


  7 in total

1.  Diffusion, mixing, and associated dye effects in DNA-microarray hybridizations.

Authors:  Jacob R Borden; Carlos J Paredes; Eleftherios Terry Papoutsakis
Journal:  Biophys J       Date:  2005-08-12       Impact factor: 4.033

2.  Two-photon excited fluorescence energy transfer: a study based on oligonucleotide rulers.

Authors:  Rina Wahlroos; Juha Toivonen; Marko Tirri; Pekka Hänninen
Journal:  J Fluoresc       Date:  2006-05-16       Impact factor: 2.217

3.  Hmx3a Has Essential Functions in Zebrafish Spinal Cord, Ear and Lateral Line Development.

Authors:  Samantha J England; Gustavo A Cerda; Angelica Kowalchuk; Taylor Sorice; Ginny Grieb; Katharine E Lewis
Journal:  Genetics       Date:  2020-10-19       Impact factor: 4.562

4.  Ultrasensitive quantification of tumor mRNAs in extracellular vesicles with an integrated microfluidic digital analysis chip.

Authors:  Peng Zhang; Jennifer Crow; Divya Lella; Xin Zhou; Glenson Samuel; Andrew K Godwin; Yong Zeng
Journal:  Lab Chip       Date:  2018-12-04       Impact factor: 6.799

5.  Detection of foot and mouth disease and porcine reproductive and respiratory syndrome viral genes using microarray chip.

Authors:  Y-C Liu; G S Huang; M-C Wu; M-Y Hong; K-P Hsiung
Journal:  Vet Res Commun       Date:  2006-02       Impact factor: 2.459

6.  Experimental optimization of probe length to increase the sequence specificity of high-density oligonucleotide microarrays.

Authors:  Shingo Suzuki; Naoaki Ono; Chikara Furusawa; Akiko Kashiwagi; Tetsuya Yomo
Journal:  BMC Genomics       Date:  2007-10-16       Impact factor: 3.969

7.  Specifically horizontally tethered DNA probes on Au surfaces allow labelled and label-free DNA detection using SERS and electrochemically driven melting.

Authors:  E Papadopoulou; N Gale; J F Thompson; T A Fleming; T Brown; P N Bartlett
Journal:  Chem Sci       Date:  2015-10-08       Impact factor: 9.825

  7 in total

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