Difference between revisions of "Sequencing"

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= TECHNOLOGY =
|title=Sequencing Service
 
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The GTC offers Next-Generation sequencing on the Illumina's NovaSeq and Element Bioscience's AVITI platform. More details on the [[Pricing| pricing]] page.
  
CAGT has two high-throughput Solexa Genome Analyzers 2.0 (Illumina), which are currently being used for a variety of applications, including ChIP-Seq, miRNA sequencing and expression sequencing. Each next-generation sequencer can process up to 16 samples per week, with a data yield of approximately 10 million reads per sample. Read lengths are 17, 26 or 36 bases.  
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We also have Nanopore PS2 solo for long read sequencing.
  
This sequencing technology consists of binding randomly fragmented DNA to an optical flowcell . Templates are “sequenced” by incorporating fluorescently labeled nucleotides in a “Sequencing-By-Synthesis” reaction. A detailed description of this process can be found here: http://illumina.com/pages.ilmn?ID=203
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= LIBRARY PREP SERVICE =
 
The system consists of a cluster generation station and a Genome Analyzer, both of which work in concert to generate and analyze flowcells. An overview of the Illumina Genome Analyzer system can be found at the Illumina website: http://illumina.com/pages.ilmn?ID=204
 
  
Illumina currently provides reagents and support for a five major sequencing applications:
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We offer many options for library prep for a wide range of applications and input levels down to RNA from single cells. Please refer to the pricing page for more information.
  
* Genome sequencing and resequencing <br>
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= PRICING =
* Paired-end sequencing <br>
 
* Digital gene expression <br>
 
* Small RNA discovery <br>
 
* ChIP-Seq <br>
 
  
Other DNA Sequencing applications: The following applications have been published but do not yet have kits from Illumina.
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For pricing information - [[Pricing| Click Here]]
  
* Genotyping: Protocols are being developed for detection of SNPs, chromosomal rearrangements and other genotyping applications. <br>
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{{SeqSubmissionGuide}}
* Whole Transcriptome Analysis: Sequencing the entire transcriptome can reveal valuable information about exon boundaries and splice variants that may be missed by microarray analysis or Digital Gene Expression. Kits for whole transcriptome analysis are expected to be available from Illumina soon.<br>
 
  
== Protocols ==
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= DATA =
  
Protocols for all of the supported technologies can be found on the Illumina website, or by contacting Illumina at (800) 809-4566. http://illumina.com/pagesnrn.ilmn?ID=70#252
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Images acquired from the sequencer are processed through the bundled Illumina image extraction pipeline to get the sequence and quality score for each base. An in-depth QC report is included in the package.
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* QC Details - [[SequencingQC|Sequencing]]
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* Formats Details - [[SequencingFormats|Sequencing]]
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== TURNAROUND TIME ==
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2-3 weeks
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= APPLICATIONS =
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Next-Generation Sequencing can be used for a wide variety of applications including ChIP-Seq, RNA-Seq, smallRNA-seq, GWAS and much more. Visit Illumina's web site or contact us to discuss your project goals.

Latest revision as of 16:51, 10 July 2024

TECHNOLOGY

The GTC offers Next-Generation sequencing on the Illumina's NovaSeq and Element Bioscience's AVITI platform. More details on the pricing page.

We also have Nanopore PS2 solo for long read sequencing.

LIBRARY PREP SERVICE

We offer many options for library prep for a wide range of applications and input levels down to RNA from single cells. Please refer to the pricing page for more information.

PRICING

For pricing information - Click Here

SEQUENCING SUBMISSION GUIDELINES

STEP 1: REGISTRATION (FOR NEW CUSTOMERS ONLY)

  • INTERNAL CUSTOMERS: Do NOT require to register in the system

All external customers please obtain a purchase order number prior to your submission. If you need a quote, please email us at wibr-genome@wi.mit.edu or call us at 617-258-8803. Blanket purchase orders are preferred, if you plan to submit multiple projects over time.


STEP 2: PREPARATION OF SAMPLES

SUBMISSION OF SAMPLES FOR LIBRARY PREPARATION

Samples should be in a buffer that does not contain trace amounts of phenol or EDTA. We will save and return any unused input material after libraries are successfully prepared.

Below are our input requirements for Library Prep Service

Sample Type Sonication/Fragmentation* Fragment Size (bp) Input requirements (ng)** Volume (ul)
Genomic Required 200 to 400 500 to 1000 10-50
ChIP Required Variable (specify) 10 to 50 10-40
RNA (total) NA NA 100 to 1000 10-50

*Unless special arrangements are made with us in advance, we will assume that your genomic DNA sample has been fragmented, by sonication or otherwise, prior to submission.

**We can work with less if necessary, but it is best if you elute your DNA in 50 ul or less if you believe we may need to use all of it. Please give us at least 10uL of sample to ensure we have enough for QC and prep.

SUBMISSION OF PRE-PREPARED LIBRARIES

We typically need 10-20 ul of each library at 5 ng/ul or greater. If your library is less concentrated than 5 ng/ul, we’ll quantify it to make sure we have enough to sequence.


STEP 3: ONLINE SUBMISSION OF SEQUENCING LIBRARIES

  • Please complete the online submission form (Link To Form Available Here). Questions or problems related to the submission should be directed to Sumeet Gupta at sgupta@wi.mit.edu
  • Every physical sample tube should have its own Genome Tech Core ID (GTC ID) number (automatically generated and emailed to you immediately after submission of the online form), even if you plan to multiplex them.
  • Please write your name GTC ID numbers on the tubes. We prefer 1.5 ml tubes.


STEP 4: DROP-OFF OR SHIPPING

When you are ready, you may hand deliver or ship samples them on dry ice to:

Whitehead Institute Genome Technology Core
455 Main Street, Room 325
Cambridge, MA 02142

Please do not ship without first contacting someone in the lab, to make sure that we are here to receive your package. Do not rely on voicemail. Please wait until you hear back from someone.

DATA

Images acquired from the sequencer are processed through the bundled Illumina image extraction pipeline to get the sequence and quality score for each base. An in-depth QC report is included in the package.

TURNAROUND TIME

2-3 weeks

APPLICATIONS

Next-Generation Sequencing can be used for a wide variety of applications including ChIP-Seq, RNA-Seq, smallRNA-seq, GWAS and much more. Visit Illumina's web site or contact us to discuss your project goals.