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| == Singular Sequencing ==
| | #REDIRECT [[BioMicroCenter:ShortRead]] |
| The Center currently hosts a G4 platform by [https://singulargenomics.com/| Singular Genomics]. Singular’s G4 platform is a short-read mid-throughput sequencing platform delivering adequate sequencing data through its proprietary sequencing-by-synthesis chemistry at a significantly competitive price point.
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| {|
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| {| class="wikitable" border=1
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| !width=100| Service
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| !width=250| Singular Sequencing
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| |-
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| |INPUT || Singular libraries
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| |-
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| |MIN VOLUME || 12 uL
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| |-
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| |MIN CONCENTRATION || 4 nM
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| |-
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| |INCLUDED SERVICES
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| * Quality Control:Fragment Analyzer and qPCR
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| * Singular Sequencing
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| * Demultiplexing
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| |-
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| |ADDITIONAL SERVICES ||
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| * Quality Control
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| * Singular library preparation
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| |-
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| |DATA FORMATS
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| *FASTQ (stored 1 year)
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| |-
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| |QUALITY CONTROL
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| * [https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ FASTQC]
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| * Basic run metrics (alignment rate, complexity)
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| * Basic RNAseq metrics (where applicable)
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| * Basic paired end metrics (where applicable)
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| * Contamination checks
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| |-
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| |PRICING || [[BioMicroCenter:Pricing#SINGULAR_G4|LINK]]
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| |-
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| |SUBMISSION
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| * MIT - [https://mit.ilabsolutions.com/service_item/new/3381?spt_id=3863 ilabs]
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| |-
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| |}
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| '''Minimum reads per lane are guaranteed if:
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| *the BMC has performed quality control,
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| *the samples are high-complexity, especially for the first 10 nt of read 1, and
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| *submitted libraries are at least 2 nM.'''
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| === Workflow ===
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| Library prep for [https://openwetware.org/wiki/BioMicroCenter:DNA_LIB DNA] or [https://openwetware.org/wiki/BioMicroCenter:RNA_LIB RNA] are available for Singular sequencing. <br> Singular library fragment size is verified via [https://openwetware.org/wiki/BioMicroCenter:QC AATI's Fragment Analyzer]. <br> Singular library is quantified via qPCR.
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| | |
| ===Requirements===
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| <OL>
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| <LI>All 4 bases (A, G, C, T) must appear within the first 10 cycles of read 1.
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| <LI>Lane-by-lane requests must be 50 or 150 paired-end and compatible with TruSeq, Nextera, TruSeq, smRNA and Solexa sequencing primers. All other requests will require a 4 lane minimum since all lanes use the flowcell's sequencing primers and this includes Illumina libraries which have been converted.
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| <LI>For standard lane-by-lane requests, the first index will be read off the S1 region of the final library, and index 2 will be read off the S2 region. Compared to a typical Illumina library, a Singular index 1 would be the reverse compliment of the i5 index, whereas index 2 would be the reverse compliment of the i7 index.
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| |}
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| == The G4 Platform ==
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| {| class="wikitable" border=1
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| !width=100| SPEC
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| !width=250| G4
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| |-
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| |'''SEQUENCER'''||[[Image:2023_Singular_G4.jpg|center|200px]]
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| |-
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| | '''READS/LANE'''<BR> Low number is minimum per lane for standard libraries||
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| * F3 per Lane: 50-100 M
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| |-
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| |'''RUN TIME'''||
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| * 50 PE 11 hours
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| * 150 PE 24 hours
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| |-
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| |'''KITS AVAILABLE'''||
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| * 100nt
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| * 300nt
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| |-
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| |'''KNOWN APPLICATIONS'''||
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| * RNAseq including single cell
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| * Gene Expression
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| * Exome
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| * Target Enrichment
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| * Metagenomics
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| |-
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| |'''KEY NOTES'''||
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| * 4-color chemistry
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| * Patterned flow cell
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| * 4-lanes per flow cell
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| * 4-flow cells per run
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| * Index replacement or anchor expansion of Illumina libraries
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| |-
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| |'''THANKS TO'''||
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| * Singular Genomics
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| |}
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