Vironomics Core Next Generation Sequencing
We can sequence a wide variety of organisms, we are not limited to viruses. The sequencers in the core are ideal for longer DNA fragment sequencing. The Ion Torrent PGM is capable of 200-400bp read lengths. The Ion Torrent S5 has the ability to increase that to 600bpª and increases the number of reads/depth available. Many clients use the longer reads for large genomes, de novo sequencing, 16S sequencing, long amplicons, etc.
Submission for one platform or the other generally depends on a) fragment length b) amount of double-stranded DNA available for sequencing and/or c) number of reads needed for your experiment.
ª 600bp costs more, please add an extra emPCR rate to the cost of the run
Examples of our seq runs and quality scores, click here
- Your custom project for Ion Torrent
- Please discuss with core manager details and costs
- Next Generation Sequencing: Cell Line verification
- NIH Grant Applications authentication of key biological and/or chemical resources
- Utilizing our Ion Torrent sequencers (more reads) allows most runs to accommodate a greater number of samples. This commercially available option, Precision ID NGS STR, allows for a plug-n-play analysis, which decreases the need for specialized bioinformatics. For more detailed information, please go to the dedicated page here.
- Plasmid Verification: We have had success with plasmid verification for various clients
- Whole Genome de novo sequencing: See our publications lists for examples
- Targeted Amplicon sequencing: custom or see off-the-shelf Ion panels
- Exome sequencing
- RNAseq (strandedness retained)
Ion Torrent PGM and S5 SUBMISSION REQUIREMENTS
They have a 97.5% per base accuracy up to 5 homopolymers.
|Submission requirements||DNA Needs Fragmentation
(gDNA, BAC, Plasmid, etc.)
|DNA Already Fragmented
double stranded cDNA, etc.)
|Total RNA†||Poly(A) RNA†||rRNA depleted RNA†|
|Purity||260/280 ratio ≥ 1.8||260/280 ratio ≥ 1.8||RIN ≥ 7||n/a||n/a|
|minimum ng/uL*||≥ 1.4||≥ 0.2||≥1.9||≥ 0.2||≥ 1.3|
|minimum total ng||50||10||100||1||10|
|ave volume to send (dependent
|40-55||10-30||≥ 10||≥ 20|
¤ Amplicons: This technology is sensitive to primer-dimers, please contact Core Manager for protocol of removing small fragments using AMPure beads. Also, if multiplexing samples, each amplicon should be similar in length.
* NanoDrop is not accurate for assessing sequencing samples. If you do need to use the NanoDrop, please send double the required input.
† Please ensure your samples do not have contaminating genomic DNA by performing a RT-minus reaction for each of your samples.
Vironomics Core Manager: 919-843-5292
On a custom basis we can make Illumina libraries and assist with submitting them to another facility. This generally requires purchase of an entire library kit since we do not keep these in stock. Please contact the Core manager to inquire about cost and staff availability.
If you have only a few samples please consider having the facility with the sequencing instrument make your libraries. Please see UNC Research Core Facilities page to explore other facilities.