Literature DB >> 12901608

A rapid method for manual or automated purification of fluorescently labeled nucleic acids for sequencing, genotyping, and microarrays.

Amy L Springer1, Lisa R Booth, Michael D Braid, Christiane M Houde, Karin A Hughes, Robert J Kaiser, Casandra Pedrak, Douglas A Spicer, Sergey Stolyar.   

Abstract

Fluorescent dyes provide specific, sensitive, and multiplexed detection of nucleic acids. To maximize sensitivity, fluorescently labeled reaction products (e.g., cycle sequencing or primer extension products) must be purified away from residual dye-labeled precursors. Successful high-throughput analyses require that this purification be reliable, rapid, and amenable to automation. Common methods for purifying reaction products involve several steps and require processes that are not easily automated. Prolinx, Inc. has devel oped RapXtract superparamagnetic separation technology affording rapid and easy-to-perform methods that yield high-quality product and are easily automated. The technology uses superparamagnetic particles that specifically remove unincorporated dye-labeled precursors. These particles are efficiently pelleted in the presence of a magnetic field, making them ideal for purification because of the rapid separations that they allow. RapXtract-purified sequencing reactions yield data with good signal and high Phred quality scores, and they work with various sequencing dye chemistries, including BigDye and near-infrared fluorescence IRDyes. RapXtract technology can also be used to purify dye primer sequencing reactions, primer extension reactions for genotyping analysis, and nucleic acid labeling reactions for microarray hybridization. The ease of use and versatility of RapXtract technology makes it a good choice for manual or automated purification of fluorescently labeled nucleic acids.

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Year:  2003        PMID: 12901608      PMCID: PMC2279886     

Source DB:  PubMed          Journal:  J Biomol Tech        ISSN: 1524-0215


  24 in total

Review 1.  Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies.

Authors:  M M Shi
Journal:  Clin Chem       Date:  2001-02       Impact factor: 8.327

2.  High-resolution semi-quantitative real-time PCR without the use of a standard curve.

Authors:  A Gentle; F Anastasopoulos; N A McBrien
Journal:  Biotechniques       Date:  2001-09       Impact factor: 1.993

Review 3.  A concise guide to cDNA microarray analysis.

Authors:  P Hegde; R Qi; K Abernathy; C Gay; S Dharap; R Gaspard; J E Hughes; E Snesrud; N Lee; J Quackenbush
Journal:  Biotechniques       Date:  2000-09       Impact factor: 1.993

4.  A DNA microarray system for analyzing complex DNA samples using two-color fluorescent probe hybridization.

Authors:  D Shalon; S J Smith; P O Brown
Journal:  Genome Res       Date:  1996-07       Impact factor: 9.043

5.  Design and synthesis of fluorescence energy transfer dye-labeled primers and their application for DNA sequencing and analysis.

Authors:  J Ju; I Kheterpal; J R Scherer; C Ruan; C W Fuller; A N Glazer; R A Mathies
Journal:  Anal Biochem       Date:  1995-10-10       Impact factor: 3.365

6.  Quantitative monitoring of gene expression patterns with a complementary DNA microarray.

Authors:  M Schena; D Shalon; R W Davis; P O Brown
Journal:  Science       Date:  1995-10-20       Impact factor: 47.728

Review 7.  Nucleic acid detection using non-radioactive labelling methods.

Authors:  E S Mansfield; J M Worley; S E McKenzie; S Surrey; E Rappaport; P Fortina
Journal:  Mol Cell Probes       Date:  1995-06       Impact factor: 2.365

Review 8.  Non-radioactive analysis of biomolecules.

Authors:  C Kessler
Journal:  J Biotechnol       Date:  1994-06-30       Impact factor: 3.307

9.  Rapid purification of fluorescent dye-labeled products in a 96-well format for high-throughput automated DNA sequencing.

Authors:  K Krakowski; J Bunville; J Seto; D Baskin; D Seto
Journal:  Nucleic Acids Res       Date:  1995-12-11       Impact factor: 16.971

10.  Convenient single-step, one tube purification of PCR products for direct sequencing.

Authors:  E Werle; C Schneider; M Renner; M Völker; W Fiehn
Journal:  Nucleic Acids Res       Date:  1994-10-11       Impact factor: 16.971

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  1 in total

Review 1.  DNA microarrays--techniques and applications in microbial systems.

Authors:  T Majtán; G Bukovská; J Timko
Journal:  Folia Microbiol (Praha)       Date:  2004       Impact factor: 2.099

  1 in total

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