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High Throughput Sequencing Facility

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Chapel Hill, North Carolina, US

About High Throughput Sequencing Facility

The HTSF is part of the Carolina Center for Genome Sciences (CCGS) and was established in 2008 with funds from the University Cancer Research Fund (UCRF) and the Lineberger Comprehensive Cancer Center (LCCC).

Our Services (8)


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Ion Torrent Sequencing

Ion Torrent Next Generation Sequencing
Price on request

The HTSF's Ion Torrent Sequencer provides excellent "sequencing by synthesis" capabilities, yielding data quickly and affordably.

The 2.5 day submission-to-data turnover time consists of:

  • library preparation (1 day)
  • emulsion PCR (1 day)
  • sequencing (1/2 day)

Currently, we offer runs with Ion 316, 318, and Proton 1... Show more »

The HTSF's Ion Torrent Sequencer provides excellent "sequencing by synthesis" capabilities, yielding data quickly and affordably.

The 2.5 day submission-to-data turnover time consists of:

  • library preparation (1 day)
  • emulsion PCR (1 day)
  • sequencing (1/2 day)

Currently, we offer runs with Ion 316, 318, and Proton 1 Chips:

Chip.........316.........318.........Proton 1
data/run...200 Mb...500 Mb...6 Gb
bp............400..........400.........200
reads.......2 M..........5 M.........33 M

Ion 314 Chip - cost per run $433 (includes reagent cost)

Ion 316 Chip - cost per run $633 (includes reagent cost)

Ion 318 Chip - cost per run $833 (includes reagent cost)

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SMRT Sequencing

Single Molecule Real-Time Sequencing
Price on request

The PacBio RS system can be used for de novo assembly, targeted sequencing, and base modification detection. There are two main sequencing options: Continuous Long Reads (CLRs) and Circular Consensus (CCS). Multiple size options can be used with these sequencing options.

We can prepare libraries of sizes; 500bp, 1kb, 2kb,... Show more »

The PacBio RS system can be used for de novo assembly, targeted sequencing, and base modification detection. There are two main sequencing options: Continuous Long Reads (CLRs) and Circular Consensus (CCS). Multiple size options can be used with these sequencing options.

We can prepare libraries of sizes; 500bp, 1kb, 2kb, 5kb, 10kb as well as amplicons without shearing. Longer inserts (10kb) are appropriate for CLR and short inserts (up to 3kb) are appropriate for CCS. Additionally CLR is appropriate for de novo assembly, while CCS is appropriate for targeted sequencing.

There are also movie sequencing options associated with the read types:

  • One 120-minute movie will maximize the read length but only capture half (~30,000 reads) of the ZMWs (reaction wells) on a SMRTcell (where sequencing takes place). This is recommended for CLR with a maximum read length > 20kb (average 4kb).

  • Two 55-minute movies will have a greater number of shorter reads (~60,000 reads). This is recommended for CCS with a maximum read length of > 9kb (average 3kb).

These standard protocols can be modified upon request.


Output per SMRTcell:

...........................CLR.................CCS
Application.........de novo............targeted sequencing
Insert sizes.........10kb................500bp or 1kb
Movie Set-up......1x120min........2x55min

Reads..............~30,000..........~60,000

Read length........max. >20kb.....max. 6kb
...........................average 4kb ...average 4kb
Output................100Mb.............30Mb
Error Rate..........15%.................2%

The number of SMRTcells you’ll need to run depends on the application. For more information please discuss your project with a HTSF PacBio tech.


Sample Requirements & Guidelines:

These are general instructions (all samples).

**All samples submitted are assumed to be measured by a flourometer (e.g. Qubit). However, if sample concentrations have been measured using a spectrophotometer (e.g. Nanodrop) please compensate for overestimation and double the input amount.

Total Genomic DNA:

Desired insert size.....DNA........Total volume
500 bp.......................500 ng.....100 µL
1 kb...........................500 ng.....100 µL
2 kb...........................1 µg.........100 µL
5 kb...........................2 µg.........150 µL
10 kb.........................5 µg.........150 µL

Amplicon:

Amplicon size*...........DNA.........Total volume
~250 bp.....................250 ng......20 µL
~500 bp.....................500 ng......20 µL
~1 kb.........................500 ng......20 µL
~2 kb.........................500 ng......20 µL
~5 kb.........................2 µg..........20 µL
~10 kb.......................5 µg..........20 µL

*Please include information on the size of the material being submitted.

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PacBio RS II

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DNA Sequencing

Price on request
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DNA Services

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Next Generation Sequencing (NGS)

Price on request
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Biology

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Biochemistry & Molecular Biology

Price on request
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Nucleic Acid Services

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