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Product Description: The DMG Diversity Navigator™ for CYP3A5 contains optimized primers and reagents that produce amplicons and sequence data specific for each exon and its flanking intronic segments. Sequence coverage is in both forward and reverse directions to assist in accurate SNP and mutation detection. Reagents are provided for analysis of 200 DNA samples.
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Component
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Amount
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Amplification primer pairs (red caps)
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11 X 100 µl
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20 µM each in 10 mM Tris HCl pH 8.0, 1 mM EDTA, 2% acetonitrile (See Table 2)
200 µl is provided for primer pair 3A5 7 A-D A
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5X PCR Buffer
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9.6 ml
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50 mM Tris HCl pH 8.3, 250 mM KCl, 10 mM MgCl2, 10% DMSO, 25 mM DTT, 1 mM each dNTP
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Sequence primers (blue caps)
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36 X 100 µl
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3 µM in 10 mM Tris HCl pH 8.0, 1 mM EDTA, 2% acetonitrile (See Table 5)
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5X Sequence Buffer
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14.4 ml
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300 mM Tris HCl pH 9.0, 7.5 mM MgCl2
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Control DNA sample
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360 µl
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2 ng/µl in 1 mM Tris HCl pH 8.0, 0.1 mM EDTA, 0.01% Tween-20
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BioMarker® EXT DNA Marker
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250 µl
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10 ng/band/µl in 10 mM Tris HCl pH 7.5, 50 mM KCl, 5 mM EDTA, 0.01% sodium azide
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BioTracker® 6X Tracking Dye
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250 µl
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30% glycerol, 0.25% bromophenol blue, 0.75% lauryl sulfate, 10 mM Tris HCl pH 8.0, 1 mM EDTA
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Reference sequences
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Facilitate sequence read assembly into contigs and alignment of polymorphic sites to other sequences. These sequences are also available for download online
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Mirror images of the reference sequences
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Reflect the biallelic alternative to the reference genotype
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Control DNA genotype
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Provides allele calls at known polymorphic sites. Control DNA Genotype is also available.
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Necessary reagents not provided:
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Purified human DNA samples, thermostable DNA polymerase (for PCR), electrophoresis reagents, additional running dye and size markers (only
samples are provided), reagents for post-PCR reaction cleanup (e.g. Exo I-SAP and BSA), dye terminator mix for dideoxy sequencing, reagents for post-sequencing reaction cleanup (e.g. SAP, 1-butanol, Sephadex™ G-50), sterile distilled water, glycerol
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Necessary equipment:
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Thermal cycler, centrifuge, electrophoresis apparatus, SpeedVac™ or incubator, sequencing instrument, PCR grade 96-well plates with seals or 200 µl PCR reaction tubes with lids, appropriate size tubes for master mix preparation
Individual components of the kit are available for purchase to suit the particular needs
of each customer. Genotyping or mutation discovery by the methods described here
are available as a contracted service from BioVentures, Inc. For more information contact us at
support@bioventures.com.
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