Our technologies support a wide range of genomic applications.

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About

Illumina Next Generation Sequencing (NGS) is a massively parallel DNA sequencing technique, using sequencing-by-synthesis chemistry to sequence millions or billions of DNA fragments simultaneously. The basic procedure involves:

  1. Library preparation: DNA or RNA-derived cDNA is fragmented into smaller pieces and special Illumina adapters are added to the blunt ends of the fragments. During this step, unique DNA barcodes carried by the Illumina adapters are incorporated into the fragments in a sample, allowing multiple libraries to be multiplexed into a single sequencing run and distinguished during data analysis.
  2. Clustering: the pool of libraries are delivered to a flow cell - a glass slide with hollow channels that capture and fix the library fragments to the surface through the Illumina adapters. Library fragments undergo clonal amplification to produce clusters, where each fragment in a cluster is identical as they originate from an original template fragment.
  3. Sequencing: nucleotides carrying a fluorophore and reversible terminator are delivered to the flow cell and incorporated into the clusters by a polymerase. The terminators inhibit extension, enabling fluorophore excitation and imaging, one nucleotide at a time. The terminators are removed, the next nucleotide is incorporated, and so on. Imaging intensities are extracted and bases are called and converted into raw basecall files.
  4. Analysis: sequencing data is demultiplexed according to the library barcodes added during library preparation and converted into FASTQ files, which can then be used as input for bioinformatic analysis. 

Applications

How it works

Illumina sequencing outputs are reported in units of megabases (Mb), gigabases (Gb) and terabases (Tb), as well as the total number of reads generated in millions or billions. Sequencing runs on Illumina instruments can take the form of single-end reads, meaning all reads are performed on the forward strand of the DNA templates - this is called Read 1. They can also take the form of paired-end reads, where the complementary strand is resynthesised after Read 1 and the forward template strand is washed away, and then reads are performed on the complementary strand in the opposite direction - this is called Read 2.

A single-end sequencing run, with read lengths of 100 bp for example, is denoted as 1 × 100 bp, and a paired-end run is denoted 2 × 100 bp. If you wish to sequence with 100 bp single-end reads, then we will use a 2 × 50 bp sequencing kit and set up the run using all 100 cycles for Read 1. Some Illumina instruments can produce paired-end reads up to 300 bp (2 × 300 bp). However, not all flow cells are compatible with all read lengths, for example, a MiSeq v2 flow cell supports read lengths up to 250 bp, but not 300 bp. The tables below detail flow cell outputs with their associated read lengths for each sequencer.

Products

ATAC-seq is a molecular biology technique used to analyse chromatin accessibility on a genome-wide scale, providing insight into regions accessible for transcription factors and regulatory proteins, active regulatory elements like promoters, enhancers, and regions of active transcription.

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We offer various sequencing solutions for your DNA-seq project, including sequencing libraries prepared by you, library preparation by us from samples you have submitted, and modifying existing protocols with flexible criteria for submitting unconventional or difficult sample types.

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HiC is a powerful molecular technique that allows scientists to study the three-dimensional organization of the genome by capturing interactions between distant DNA segments. With HiC, researchers can gain insights into the spatial arrangement of chromosomes and how it influences gene regulation and genome function.

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Uncover the DNA methylation status of your samples with methyl-seq: a cutting-edge epigenetic profiling technique that resolves DNA methylation status through sequencing, providing valuable insights into their functional implications in various biological processes.

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We provide RNA-seq library preparation for a wide range of sample types and project goals, including strand-specific mRNA prep for transcriptome sequencing or total RNA with ribo-depletion for the preservation of non-coding RNA, as well as low input, degraded and small RNA library prep.

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Substitute bulk RNA-seq with single-cell sequencing for the resolution of cell populations, differential transcriptomics, V(D)J immune profiling and fixed RNA profiling. We use 10x Genomics Chromium iX for cell barcoding and construction of various library types, sequenced on Illumina platforms.

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Equipment

The MiSeq is a versatile and cost-effective benchtop sequencer, supporting targeted resequencing and small-genome sequencing, as well as library QC for larger sequencing projects.

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The NextSeq 2000 is a flexible and efficient benchtop sequencer that utilises super-resolution optics and enhanced patterned flow cell technology to maximise the output of the sequencing run.

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The NovaSeq 6000 is a powerful high-throughput sequencer, offering scalability and flexible output options to suit your research goals. Dual flow cell compatibility enables simultaneous sequencing runs and flow cell design allows segregation of libraries into separate lanes.

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The NovaSeq X Plus is Illumina's latest technological innovation, combining dual flow cell configuration with the most advanced and efficient XLEAP-SBS chemistry to generate massive quantities of data with unprecedented quality.

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Cost

Please contact us to enquire about price.

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How to order

Order forms for the following Illumina services can be downloaded. Options:

  1. BRF prepares library on behalf of the customer
  2. The customer prepares the library.

Terms & conditions

General

  1. The BRF is an NCRIS Bioplatforms Australia (BPA) genomics node, supported by BPA, ANU and JCSMR. The BRF is a core facility housed within JCSMR and does not endorse any samples that it processes. The Clients for which the BRF performs services are not permitted to use the name or logo of the ANU or BRF facility in any marketing, advertising or promotional material.
  2. No samples or orders will be processed without a sample submission/order form and valid charge code, authorised by the signature of the PI/Lab Head, or by granting of electronic access to BRF ordering systems with the authority of the PI/Lab Head.
  3. All work performed in the BRF, whether full service or not, and the use of any BRF resources should be acknowledged in all publications arising from that work. This should be in the “Material and Methods” section of the paper (see examples below). Any further individual acknowledgements are solely at your discretion.
  4. A reference to the publication should be sent to the Manager of the BRF once the paper is published (includes theses).
  5. BRF collaborations and co-authorship: although the BRF is primarily a service unit, there are instances where BRF staff make significant contributions to either the technical or intellectual input of the project. This should be discussed with the staff member concerned prior to the commencement of the collaboration.
  6. The BRF retains intellectual property rights generated in the processing of samples.
  7. The Client is responsible for ensuring they have secured the necessary ethics approvals prior to requesting services from the BRF.
  8. Libraries and other materials generated by the BRF are the property of the BRF and can only be sequenced or manipulated on instrumentation owned by the BRF. The BRF reserves the right to refuse handover of BRF-owned samples and material for the purpose of manipulation or sequencing on third party instrumentation.
  9. The BRF may require the Client to provide it personal identification information for the purpose of contacting the Client. This information includes, but is not limited to, email address, workplace phone number, institution address, laboratory group leader(s) and their contact information etc.

 

Services & Payment

  1. Unless specified, amounts indicated in quotations and correspondence are exclusive of GST.
  2. The BRF is not responsible for importation fees and customs clearance fees for samples/libraries being submitted by the Client to the BRF.
  3. ANU Clients must supply a valid ANU charge code in BRF submission documents for the BRF to proceed with services. The BRF reserves the right to delay services and NGS data handover until a valid ANU charge code is supplied to the BRF.
  4. For Clients external to the ANU, services provided by the BRF that amount to less than the credit card limit within the Clients’ institutional guidelines can be paid for by a credit card. The Client agrees to provide the BRF with a submission document containing their contact information for the issuing of an invoice.
  5. Client contact information will be used solely for communication with the BRF regarding services and projects and will not be distributed to third parties.
  6. For Clients external to the ANU requesting services that amount to more than the credit card limit of the Clients’ institutional guidelines, the BRF will issue a quote to the Client for the agreed services.
  7. If a Client external to the ANU indicates the use of an external payment portal that charges the ANU a percentage of the invoice amount, this additional amount will be included as a surcharge on the invoice issued to the Client. This term will be indicated on the quote issued to the Client for raising a purchase order.
  8. The Client must use this quote to raise a purchase order with the finance department of their institution/organisation, and to provide the BRF with a digital copy of the purchase order.
  9. The BRF reserves the right to delay commencement of services or handover of NGS data until a purchase order has been submitted.
  10. The Client agrees to paying for all library preparations regardless of their outcome.
  11. The BRF agrees to perform the services for you (the Client) with due care and skill, and to include all applicable quality control checks throughout sample processing to ensure manufacturer’s specifications are met. For samples that meet the quality and quantity requirements as outlined in the Illumina Sample Submission Guidelines document, the Client is guaranteed the data output and quality outlined in Illumina instrument specifications. The Client agrees that the BRF is not responsible for suboptimal data output/quality and cannot guarantee optimal data from:
    • samples that do not meet BRF quality control checks as outlined in the Illumina Sample Submission Guidelines document.
    • libraries prepared by the Client that do not reflect BRF quality controls as outlined in the Illumina Sample Submission Guidelines document, or that do not reflect Illumina recommendations.
    • libraries derived from assays that deviate significantly from recommended manufacturer’s protocols, such that the BRF suspects the sequencing performance will be impacted as a result of these deviations.
    • samples/libraries that have been improperly handled or stored.
  12. Turnaround times indicated verbally or in writing by BRF staff are on a ‘reasonable endeavours’ basis and not a binding commitment.
  13. The BRF reserves the right to refuse any requests for Service at our discretion. In the event that the BRF is unable to or rejects providing a service, the BRF will notify the Client promptly.
  14. The Client acknowledges that Illumina data generated by the BRF is for research use only and will not be used as part of any clinical, diagnostic or therapeutic procedures, unless performed under the guidelines of a workflow established by an accredited entity that permits the BRF to perform such services.
  15. Illumina NGS data generated by the BRF can only be used for the Clients’ research project and cannot be sold or distributed to third parties, unless prior written consent from the BRF is obtained.
  16. The BRF reserves the right to review service costs if the number of samples submitted changes, or if the samples are provided in smaller batches than specified in the quotation. 

 

Samples

  1. The BRF will advise against preparation of samples that deviate too heavily from the requirements outlined in the Illumina Sample Submission Guidelines document and will proceed only if the Client provides written permission for the BRF to perform the services.
  2. The BRF reserves the right to dispose of remaining sample/library material (if any) 1 month after handover of NGS data.
  3. The Client agrees that upon submission of samples, the BRF is afforded a non-transferable license to use these samples to fulfil the agreed services owns all intermediate and all products derived during the processing of samples including libraries and PCR-amplified nucleic acids.  
  4. The Client agrees to cover the cost of any consumables ordered on their behalf if the Client fails to supply the correct quantity and quality of the starting material required for processing.
  5. The Client must supply safety data sheets (SDS) for hazardous samples, reagents and materials upon submission to the BRF.
  6. It is the responsibility of the Client to secure sufficient insurance for the transportation of samples/libraries to the BRF. The BRF assumes no responsibility for the loss of samples/libraries during transportation, and assumes no responsibility for mishandling or damage to samples/libraries prior to submission.
  7. The BRF assumes no responsibility for the misplacement, mishandling, damage or loss of samples that are left in an unattended location by the Client if delivering in-person. The Client agrees to handover samples/libraries directly to a BRF staff member during office hours or an agreed appointment time.
  8. The Client must ensure the samples/libraries submitted to the BRF are de-identified and do not contain or display confidential personal information, unless necessary privacy consent is provided to the Client and the BRF.

Indemnity

  1. The BRF is indemnified for the Clients’ loss, damage, claims or expenses incurred as a result of or in association with:
    1. a breach of these terms and conditions by the Client
    2. a negligent or unlawful action by the Client
    3. loss of samples due to lack of insurance, improper shipping conditions, handling, customs delays, incorrect delivery address or delivery outside of working hours or appointment times. 

 

Data Retention

  1. Clients are responsible for archiving and safely storing NGS data generated by the BRF. Illumina NGS data will, upon confirmation of reception, immediately become the responsibility of the Client. The Client agrees to notify the BRF that Illumina NGS data has been received without errors within 1 week of data handover if electronically transferred.
  2. The BRF will retain Illumina NGS data for 3 weeks, after which the BRF will delete demultiplexed data and archive raw NGS data for 6 months. The BRF reserves the right to refuse requests to retrieve and re-analyse raw NGS data, except under circumstances that will be assessed at our discretion.
  3. The BRF reserves the right to permanently delete data 6 months after being archived if circumstances require.
  4. The BRF will not be liable to the Client for any loss, claim or demand made by the Client due to improper handling, use, storage or deletion of data by the Client.

 

 

Acknowledgement in Publications

  1. Although the BRF is primarily a service unit, there are instances where BRF staff make significant contributions to either the technical or intellectual input of the project. Acknowledgement of individual BRF staff as collaborators or co-authors are solely at the Client’s discretion, but we encourage the client to discuss this with the BRF at the start of the project.
  2. The client agrees to share a reference any publication, including theses, with the Manager of the BRF once the paper is published. Contact details can be found in the BRF’s website.
  3. All work performed in the BRF, as well as the use of any BRF resources must be acknowledged in all publications and presentations arising from that work, both in the Methods and in the Acknowledgement sections. Mentions and acknowledgments shall also state that NCRIS-enabled Bioplatforms Australia infrastructure were used for the work.

 

Examples

  1. "Mouse gDNA samples were submitted for library preparation using Illumina DNA Prep, Tagmentation and sequenced on the Illumina NovaSeq X Series. This work was performed by the BRF – an NCRIS supported Bioplatforms Australia genomics node at The John Curtin School of Medical Research, ANU.”
  2. "Human PBMC samples were submitted to the BRF - an NCRIS supported Bioplatforms Australia genomics node at The John Curtin School of Medical Research, ANU. The BRF generated 10x Genomics Chromium 3' Single-Cell Gene Expression libraries with Feature Barcoding and sequenced on the Illumina NovaSeq 6000, analysed by the BRF using 10x Genomics cellranger v7.2.0 analysis pipelines."
  3. "We prepared sequencing-ready amplicon libraries for Illumina sequencing to the BRF – an NCRIS supported Bioplatforms Australia genomics node at The John Curtin School of Medical Research, ANU. The work carried out by the BRF included library pool QC and sequencing on the Illumina MiSeq with a 2 × 250 bp read length configuration.”