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Pyrosequencing
 

OVERVIEW
DESCRIPTION  

Pyrosequencing is a simple and robust technique that sequences by synthesis. Using a short PCR product as a template, bases are added systematically and incorporated into a replicating DNA strand. Pyrosequencing utilizes a series of enzymatic reactions to detect pyrophosphate that is released during base incorporation and uses the detection data to establish the sequence of the template strand.


Click here for animation of this process

Pyrosequencing data is a quantitative measurement of each detected nucleotide and is ideal for measuring relative amounts of CpG methylation, allele quantification, and mutation analysis. Pyrosequencing ensures precision and accuracy, reliable detection and quantification of sequence variation, down to at least the 5% mutation level.

INSTRUMENT
Qiagen PyroMark Q24 - Pyrosequencing  technology for real-time, sequence-based detection for quantification of sequence variants (SNPs/mutation detection) )and epigenetic methylation (CpG analysis). For LUMA analysis please contact us.

SERVICE OPTIONS
Our service can be tailored to individual requirements and may include all or a combination of the following:

• Assay design
• Oligonucleotide synthesis
• Assay optimization
• Sample assay
• Interpretation and troubleshooting of results

Please contact us to discuss your individual requirements.

READ LENGTHS
Typically 50-60 bases, but, as with any sequencing technology, the maximum length will be dependent on the template secondary structure, base content, quality of PCR-product and other parameters.

TURN-AROUND TIME
Data should be available within 2-3 business days of delivery.

Contact:   Alberto Lovell
Phone: (650) 723-3189
Email: alberto5@stanford.edu