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SNaPshot™ Multiplex Kit (Applied Biosystems™)

The SNaPshot® Multiplex Kit is part of the SNaPshot® Multiplex System. The SNaPshot® Multiplex System primer extension-based method enables multiplexing up to 10 single nucleotide polymorphisms (SNPs). Use this system to screen and confirm SNPs, detect minor sequence variations, assess DNA methylation, fingerprint bacterial artificial chromosomes (BACs) or general labeling of restriction fragments.

• Interrogate up to 10 SNPs from different amplicons in a single base extension reaction
• Separate SNP loci that differ by a single base pair
• Increased sensitivity compared to standard sequencing allows detection of minor variants (e.g. somatic mutations) in a test sample
• Reduce the complexity of your experiments with this easy to use and cost-efficient genotyping system
• A large body of scientifc literature demonstrates the utility for genotyping, mutation detection and mapping

The SNaPshot® Multiplex Kit is a part of the SNaPshot® Multiplex System:
1) SNaPshot® Multiplex Kit (100, 1000, 5000 reaction sizes)
2) GeneScan™-120 LIZ™ Size Standard
3) Matrix Standard Set DS-02 for dye set E5 appropriate for your instrument

Each of the three components above is required to run the SNaPshot® Multiplex Kit. The Matrix Standard set, however, is only required at a one-time initial set up to calibrate the instrument for the SNaPshot-specific dye set E5.

4) GeneMapper® Software v.4.0 (or higher version) is the recommended secondary data analysis software that enables set up of user-defined panels and bins for automated data analysis.

Alternatively, simple, manual peak analysis is possible using PeakScanner™ 1.0 software (free download from website) for the 310, 3100. 3130, and 3730 instruments (PeakScanner is not compatible with data from 3500 instruments). For the 3500 instruments the primary data analysis software on the instrument allows fragment sizing and color separation similar to PeakScanner.

5) The SNaPshot primer Focus Kit (4329538) is a useful accessory kit for single experiment or assay (panel) design: Analyze individual primers for their approximate sizing location. This will determine whether or not two oligonucleotides will produce overlapping signals when run simultaneously and enable the setting of proper loci windows (bins) for GeneMapper analysis software.

Screen and Confirm SNPs
The SNaPshot® Multiplex System is the perfect tool for SNP screening and validation. The kit offers a one-tube single-base extension⁄termination reagent to label DNA fragments. Run DNA fragments on any Applied Biosystems capillary electrophoresis instrument with the GeneScan™-120 LIZ® Size Standard to indicate the size of labeled fragments. Then use GeneMapper® Software to analyze the data and generate allele calls. The SNaPshot® Multiplex Kit allows multiplexing during single-base extension of up to 10 primer-template combinations in a single tube-single capillary format.

Assess DNA Methylation
The study of methylation⁄epigenetics is emerging as an important component of cancer research. In a typical assay to detect methylation, bisulfite treatment of DNA deaminates non-methylated cytosine and converts it to uracil while methylated cytosine remains unreactive. The subsequent step of PCR amplification coverts uracil bases to thymine. Use SNaPshot® todetect the base differences in treated and untreated samples to learn the methylation status of your samples.

Fingerprint BACs
New BAC libraries require a rapid, efficient method for characterization (also called fingerprinting) and assembly of clones into contigs (contiguous consensus sequences), which are then arrayed into physical maps of the chromosome. Use the SNaPshot® Multiplex Kit to fingerprint clones by labeling BAC fragments after digestion with restriction endonucleases. The labeled fragments can then be separated and detected on any Applied Biosystems capillary electrophoresis instrument. Sizing information from GeneMapper® Software v 4.0 or higher is imported into subsequent editing and contig assembly programs.

In a similar fashion, any restriction fragment with a 5' overhang can be labeled with a fluorescent dye using the SNaPshot reagent.

For Research Use Only. Not for use in diagnostics procedures.

PureLink™ HiPure BAC Buffer Kit (Invitrogen™)

The PureLink® HiPure BAC Buffer Kit provides additional buffers and reagents for performing BAC (Bacterial Artificial Chromosome) DNA isolation using the PureLink® HiPure Plasmid Kits. The PureLink® HiPure Plasmid Kits utilize a proprietary anion-exchange resin which allows you to purify high-quality BAC DNA from E. coli cultures. BAC DNA requires the use of large bacterial culture volumes to obtain a reasonable DNA yield. The additional buffers and reagents needed for working with larger bacterial cultures are provided in this Buffer Kit. From a 100 ml culture, you can expect to purify ~40 µg of BAC DNA. The PureLink® HiPure BAC Buffer Kit provides reagents sufficient for five BAC Maxipreps, or 25 BAC Midipreps, or 100 BAC Minipreps.

PureLink™ HiPure Plasmid Midiprep Kit (Invitrogen™)

The PureLink® HiPure Plasmid Midiprep Kit is designed to isolate transfection-grade plasmid DNA from E. coli. The Midiprep Kit protocol will typically yield 100–350 µg plasmid DNA from 15–25 mL bacterial culture, at a purity that is comparable to that achieved by two passes through cesium chloride gradients. Advantages of the PureLink® HiPure Plasmid Midiprep Kit include:

Labor and time saving—no need for extra steps to remove endotoxin and other contaminants
High-purity, low-endotoxin product—pure enough for mammalian cell transfection
Versatility—purify any type and size of plasmid DNA, including BAC, bacmids, and ssM13 DNAs

Anion-exchange chromatography for plasmid purification
The PureLink™ HiPure Plasmid DNA Purification Kits use a patented anion-exchange resin to purify plasmid DNA to a level equivalent to two passes through CsCl gradients. The resin combines excellent capacity with a fast flow rate, high resolution, high yield, and efficient endotoxin removal. Plasmid preparations can typically be completed in less than 2 hours.

Contaminant-free DNA
Unlike cesium chloride gradient protocols, PureLink® HiPure Plasmid Purification Systems do not use organic solvents, ethidium bromide, or cesium chloride, all of which are troublesome to work with and to dispose of. Typically, plasmid DNA prepared using the PureLink® HiPure Plasmid Midiprep Kit will have an A260/A280 ratio of >1.80, indicating that the DNA is reasonably free of proteins that might interfere with downstream applications. Endotoxin levels are typically within 0.1–1 EU/µg, making this plasmid DNA ideal for mammalian cell transfection.

3730 DNA Analyzer 48-Capillary Array, 50 cm (Applied Biosystems™)

This is a Capillary Array for the Applied Biosystems® 3730 DNA Analyzers. Each array contains 48 capillaries, 50 cm long. Used for sequencing runs.

Usage per array: 300 runs (with use of Applied Biosystems reagents).

3730 DNA Analyzer 48-Capillary Array, 36 cm (Applied Biosystems™)

This is a Capillary Array for the Applied Biosystems® 3730 DNA Analyzer. Each array contains 48 capillaries, 36 cm long. Used for both sequencing and fragment analysis runs.

Usage per array: 300 runs (with use of Applied Biosystems reagents).

3730xl DNA Analyzer 96-Capillary Array, 50 cm (Applied Biosystems™)

This is a Capillary Array for the Applied Biosystems® 3730xl DNA Analyzer. Each array contains 96 capillaries, 50 cm long. Used for standard sequencing.

Usage per array: 300 runs (with use of Applied Biosystems reagents).

3730xl DNA Analyzer 96-Capillary Array, 36 cm (Applied Biosystems™)

This is a Capillary Array for the Applied Biosystems 3730xl DNA Analyzer. Each array contains 96 capillaries, 36 cm long. Used for sequencing, microsatellite, SNP genotyping, and fragment analysis runs.

Usage per array: 300 runs (with use of Applied Biosystems reagents).