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Next Generation DNA Sequencing (NGS) and library construction

The Illumina MiSeq platform represents an in-step platform for DNA sequencing but still have the capacity to compleatly sequence small and mid-sized genomes.
Courtesy of Illumina, Inc.

Illumina MiSeq

The Illumina MiSeq System, which is a step-in platform among various Illumina sequencing platforms, will still facilitate your research with a wide range of sequencing applications. It is capable of automated paired-end reads and up to 50M reads per run, and delivering up to 600bp of sequence data per target molecule.

Applications are small genome sequencing, 16S metagenomics or any other amplicon survey, and RNA sequencing (RNA-Seq). Multiplexing and adjustable read-lengths allow flexibility. Multiplex sequencing by using barcode sequences allows large numbers of libraries to be pooled and sequenced simultaneously during a single run on the MiSeq high-throughput instrument.

We offer:

  • Construction of Illumina ready-to-sequence DNA- or cDNA-libraries, including barcoding and according to best practice
  • Sequencing, using a portfolio of various Illumina sequencing kits ranging between 2x25bp up to 2x300bp (see MiSeqSpec; .pdf; 906 kB)

Sample requirements

As always, samples of DNA or RNA should be of adequate quality. For DNA usually highly purified and high-molecular weight material is needed. For RNA we expect highly purified material of high integrity, i.e. there should be minor signs of degradation. For both DNA and RNA we offer quality measurements and advise on eventual improvements. The amounts and concentrations needed varies for applications and are shared request if you request it.

Optional aspects

To be considered:

  • Barcoding/multiplexing
  • Sequencing length, single or paired end reads
  • Number of reads

Forms

  • A Sequencing Form (see MiSeqForm; .xlsx; 30 kB) is required for each project.
  • A Sample Sheet is required for each sequencing run, which is a listing samples and barcodes used, in addition, electronically accompanying the deposition of samples. A Sample Sheet is usually constructed using a downloadable software tool provided by Illumina, the Experiment Manager (IEM) and the produced output is a file that is needed by the MiSeq sequencer.

Pricing

Costs are for either library construction or sequencing, or a combination of these two. There are multitude of kits/protocols available for library construction, and we only keep a few in stock. Costs for library construction will depend on your preferences, but usually are in the range between 500SEK up to 1500SEK per library. Besides there will be minor costs for quality assurance along the procedure of the construction.

Costs for sequencing depends on the desired read length and the number of reads. Prices are approximate and varies between 2800SEK up to 14800SEK per run (see MiSeqOptions; .pdf; 606 kB). On top of that, we add a 30% service charge (as of 2019-04) on the total to cover for other consumables and service contracts. For all users “overhead” charges areadditionally applied. For users outside Lund University we also need to charge for labor on an hourly basis.

Page Manager:
. A tiny flow cell, 1.5x3cm in size, that is representing the active surface on which approx. 20 million DNA molecules are sequenced simultanously.
Courtesy of Illumina, Inc.

Contact information

The sequencing Facility is situated at Department of Biology, in the Ecology Building, Sölvegatan 37 on Floor 2 (staircase A).

For inquiries about Illumina MiSeq sequencing, analyses on Agilent BioAnalyzer and/or fragmentation of DNA/RNA on Covaris, contact:

Tomas Johansson
Researcher
MEMEG

Telephone: +46 46-222 45 49
Mobile: +46 709-57 18 04
E-mail: Tomas [dot] Johansson [at] biol [dot] lu [dot] se

For inquiries about ABI Sanger Sequencing and Fragment Analysis contact:

Anna Sterngren
Research Engineer
MEMEG

Telephone: +46 46-222 08 70
Mobile: +46 72-203 33 56
E-mail: Anna [dot] Sterngren [at] biol [dot] lu [dot] se

Funding

The facility acknowledges generous financial support during the years from the Knut and Alice Wallenberg Foundation (SWEGENE programme), the Crafoord Foundation and the Faculty of Science, Lund University.