A rapid method for manual or automated purification of fluorescently labeled nucleic acids for sequencing, genotyping, and microarrays.
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.
Springer
Amy L
AL
Prolinx. Inc., Bothell, WA, USA. amy.springer@sbri.org
Booth
Lisa R
LR
Braid
Michael D
MD
Houde
Christiane M
CM
Hughes
Karin A
KA
Kaiser
Robert J
RJ
Pedrak
Casandra
C
Spicer
Douglas A
DA
Stolyar
Sergey
S
D001483:Base Sequence; D011232:Chemical Precipitation; D018076:DNA, Complementary / Q000302:isolation & purification; D005456:Fluorescent Dyes / Q000737:chemistry / Q000302:isolation & purification*; D020869:Gene Expression Profiling; D005838:Genotype; D006193:Haemophilus influenzae / Q000737:chemistry; D008280:Magnetics; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000737:chemistry / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis; D012329:RNA, Bacterial / Q000302:isolation & purification; D012331:RNA, Fungal / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D005456:Fluorescent Dyes; D009696:Nucleic Acids; D012329:RNA, Bacterial; D012331:RNA, Fungal