$940.00 per chip (not include tech time), two bar coded sample libraries per chip
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.
Sample Submission: login through https://client-seqcore.brcf.med.umich.edu/ then choose ‘Fragment Analysis Service’
50 to 200 ng of gDNA at a concentration of 10ng/ul.
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