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Sequencing
Sanger Sequencing of single samples usually using PCR products or plasmids as template and sequenced using automated capillary-based AB 3730 and AB 3730XL DNA Analyzers.
Pyro Sequencing for mutation, methylation studies and sequencing of short sequences about 100 bases or less using PyroMark
sequencer from Qiagen. Sequencing_Handout
Genotyping
Fluorescently labeled fragments (MS, AFLP, tRFLP, etc.) are run on an Applied Biosystems 3730 Genetic Analyzer and then the size is interpreted using the GeneMapper or GeneMarker analysis software by comparing the fragment size to the internal size standards added to the samples.
qPCR
Quantitative PCR (qPCR) is often used for quantitation of expressed genes. Detection is possible with either a fluorescently labeled oligonucleotide probe (Taqman) or non-specific DNA binding chemistry (SYBR green). These options all allow you to view data as it amplifies in real time eliminating the need for post-PCR processing.
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Plate Readers
The Molecular Dynamics plate readers use monochromactic optics to read from cuvettes or plates in three modes: UV-Vis abosorbance, Fluorecence Intensity and Glow Luminescence.
Imaging
The imaging page lists the instruments available for imaging gels,blots, etc. for luminescnence, fluorescence, IR and phosphor imaging. Instruments available are TyphoonTrio, FluorochemHD, FluorochemQ, Odyssey and NightOwl imagers.
Automation
Qiagility is useful to set up qPCR assays or traditional PCR in 96 or 384 well plates. The facility's tested assay using 2 ul gave 0.3 cv or better within replicates.
Biomeks can be used for plate replicates, assay setup, purifications or any automated pipetting needs.
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Instruments
2400 thermocycler, AB 3730, AB 3730XL, Biomek FX, Biomek NX, Flexstation 3, FluorochemHD, FluorochemQ, Fragment Analyzer, NanoDrop, NanoVue, NightOwl, Odyssey, PyroMark, Qiagility, QuantStudio, Sorvall plate centrifuge, Spectramax M3, Typhoon Trio, Veriti themocycler, and ViiA7.
