BioMicroCenter:Genome Seq: Difference between revisions

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{{BioMicroCenter}}
{{BioMicroCenter}}


== Genome Seq ==
[[Image:BioMicroCenter-72_72_errorRate.png | 400px | right]]Both the HiSeq and the Genome Analyzer support ''de novo'' sequencing with extended read lengths, high passed-filter read counts, and the option to run lanes as paired-end for additional read length and long-range sequence information. <br><br>
[[Image:BioMicroCenter-72_72_errorRate.png | 400px | right]]
''De novo'' sequencing can be done using the Genome Analyzer. Read lengths have recently been increased and data associated with these long reads look very good! http://illumina.com/downloads/ch2-ILMN_ProdGuide_DNAsequencing.pdf


== Sample Submission Guidelines ==
== Sample Submission Guidelines ==
Users may submit unprepared genomic libraries to the BioMicro Center for [[BioMicroCenter:Illumina_Library_Preparation|Sample Prep]] followed by Illumina sequencing, or they may submit already-prepared libraries directly to sequencing.<br><br>
Standard Illumina protocols for sample preparation can be found [[BioMicroCenter:Protocols| here]]. Illumina Paired End sample preparation kits can be purchased directly from Illumina or through the BioMicro Center for $3,600.00 plus shipping costs (10 sample preps per kit). To request an order or for more information please contact [[BioMicroCenter:People|Kevin Thai]]. Ready-for-sequencing samples should be gel-purified according to your protocol and contain least 5uL of solution at a concentration of at least 10 nM in EB/ Tween 0.1%. [[BioMicroCenter:Sequencing_Quality_Control|Quality control]] will be run on all submitted samples.<br><br>
Samples generated using standard kits with Illumina PCR primers can be submitted without sequencing primers. Sample preparations using customized PCR primer sequences should be submitted with a complementary sequencing primer.


'''Users are required to prepare their own libraries for sequencing''' by following the corresponding Illumina [[BioMicroCenter:Protocols| Protocols]]. If this is a service that you would like the BMC to perform in the future please visit the [[BioMicroCenter:Ideas|Ideas]] page.
[[BioMicroCenter:Forms#Downloads|'''Sample Submission Forms''']]
 
Samples which are ready for sequencing should be gel-purified according to your protocol, and provided as at least 5uL of solution at a concentration of at least 10 nM in EB/ Tween 0.1%. To calculate your sample’s molar concentration, you must know the average length of your product.
 
When submitting a sample you must supply the BMC with the molar concentration, average length, and species of origin of all samples.
 
We encourage all those submitting samples to allow us to run QC analysis, using the [[BioMicroCenter:2100BioAnalyzer|2100 BioAnalyzer]], to obtain/confirm concentration information. Cyber green and qPCR QC analysis also coming soon!
 
Samples generated using standard kits with Illumina PCR primers can be submitted without sequencing primers. Custom sample preparations using customized PCR primer sequences should be provided along with a complementary sequencing primer.
 
[[BioMicroCenter:Forms#Downloads|Sample Submission Forms]]


== Data ==
== Data ==


Images acquired from the solexa sequencer are processed through the bundled solexa inage extraction pipeline to get the sequence and quality score for each base. The data is aligned to a reference genome, if requested, using an interative ELAND algorithm. An in-depth QC report is included in the package. Sample QC report is here.
Images acquired from the solexa sequencer are processed through the bundled solexa inage extraction pipeline to get the sequence and quality score for each base. The data is aligned to a reference genome, if requested, using an interative ELAND algorithm. An in-depth QC report is included in the package.
 
== Turnaround Time ==
 
Each Genome Analyzer processes 8 samples per run, or 7 samples plus a control. The control is typically used to improve sequence quality. Full flowcells can be run usually within two weeks of submission. Partial submissions of less than eight samples (or 7 with control) are put into a project queue, where they join existing samples or await others before processing. Wait times for partial submissions vary depending on demand from other users.
 
Once the run has begun approximately 10 days are required for clustering, sequencing, and data analysis for a Paired End read (36 bp read x 2).
 
== Pricing ==
 
Illumina sequencing is currently available '''only''' for labs associated with the BioMicro Center [[BioMicroCenter:CoreDeps|Core departments]]. Sequencing is currently available at an '''introductory''' price:
      '''Paired End Reads (priced per lane):'''
            7 lane run | 8 lane run
                $1105 | $980
 
== Genome Seq Literature ==
 
<biblio>
 
#Paper1 pmid=18677319

Latest revision as of 09:39, 30 September 2011

HOME -- SEQUENCING -- LIBRARY PREP -- HIGH-THROUGHPUT -- COMPUTING -- OTHER TECHNOLOGY

Both the HiSeq and the Genome Analyzer support de novo sequencing with extended read lengths, high passed-filter read counts, and the option to run lanes as paired-end for additional read length and long-range sequence information.

Sample Submission Guidelines

Users may submit unprepared genomic libraries to the BioMicro Center for Sample Prep followed by Illumina sequencing, or they may submit already-prepared libraries directly to sequencing.

Standard Illumina protocols for sample preparation can be found here. Illumina Paired End sample preparation kits can be purchased directly from Illumina or through the BioMicro Center for $3,600.00 plus shipping costs (10 sample preps per kit). To request an order or for more information please contact Kevin Thai. Ready-for-sequencing samples should be gel-purified according to your protocol and contain least 5uL of solution at a concentration of at least 10 nM in EB/ Tween 0.1%. Quality control will be run on all submitted samples.

Samples generated using standard kits with Illumina PCR primers can be submitted without sequencing primers. Sample preparations using customized PCR primer sequences should be submitted with a complementary sequencing primer.

Sample Submission Forms

Data

Images acquired from the solexa sequencer are processed through the bundled solexa inage extraction pipeline to get the sequence and quality score for each base. The data is aligned to a reference genome, if requested, using an interative ELAND algorithm. An in-depth QC report is included in the package.