Ion Proton Exome Sequencing

This service uses the AB AmpliSeq exome enrichment system (according to manufacturer's recommended protocols) in order to generate the exome-enriched genomic library. That library is then sequenced on our Ion Proton sequencer. The Ion Proton sequencer system uses semiconductor sequencing technology as a method for sequencing DNA. Nucleotide additions are detected during synthesis of a new DNA strand through the measurement of H+(Protons), released each time an individual nucleotide is incorporated on the strand. The identity of the newly-added nucleotide is known based on which nucleotide was currently being pumped through the cell at the time.

 

Ion Proton Exome Sequencing

Platform
Ion Proton Sequencer
Category
Exome Sequencing
Primary Contact

Suzanne Genik genik@umich.edu 734-764-8067

Typical Cost Click here to view internal pricing list!
Click here to view external pricing list!
Typical Output

Raw data: 10Gb of data per run

Run time 2-4 hours

Data file formats: BAM, FASTQ, and VCF

Read length up to 200 bp,

Number of reads passing filter is 60-80 Million

Approximate average read depth:

Running one exom sample per chip average coverage about 240X,

Running two exome samples per chip average coverage about 120X,

Running three exome samples per chip average coverage about 80X,

Running two exomes per Ion PI chip, usually see an average of 20X at 95-98%.

98% of target bases are covered at 10X and 94% of targeted bases at are covered at 20X ,

The actual read depth per sample may vary as a result of barcode balance and chip loading.

How to Request
Sample Requirements

50 to 200 ng of gDNA at a concentration of 10ng/ul.

Typical Turnaround

1 to 2 weeks

More Information

The Ion Proton Sequencer uses semiconductor sequencingtechnology as a method for sequencing DNA.The synthesis of sequences are generated through themeasurement of the direct release of H+(Protons).

The semiconductor chips containing microwells with templateDNA strands. Each time a nucleotide is incorporated in thesequence it triggers the release of a hydrogen ion.The release of a hydrogen ion in the reaction changes the pHof the reaction.

A series of electrical pulses from the chip is transmits to acomputer. The pulses are transmitted into a DNA sequence.

The Ion AmpliSeq technology uses a Exome Panel Kit of 12Primer pools in the construction of the Amplicon libraries.The PCR amplification of genomic target regions produce arange of DNA fragments between 200-400bp.The library is amplified using emulsion PCR.

Website source for Ion Proton