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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. 
{| class="wikitable" border=1
  !width=100| Service
  !width=250| Singular Sequencing
  |-
  |INPUT || Singular libraries
  |-
  |MIN VOLUME || 12 uL
  |-
  |MIN CONCENTRATION || 4 nM
  |-
  |INCLUDED SERVICES
  |
* Quality Control:Fragment Analyzer and qPCR
* Singular Sequencing
* Demultiplexing
  |-
  |ADDITIONAL SERVICES ||
* Quality Control
* Singular library preparation
  |-
  |DATA FORMATS
  |
*FASTQ (stored 1 year)
  |-
  |QUALITY CONTROL
  |
* [https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ FASTQC]
* Basic run metrics (alignment rate, complexity)
* Basic RNAseq metrics (where applicable)
* Basic paired end metrics (where applicable)
* Contamination checks
  |-
  |PRICING || [[BioMicroCenter:Pricing#SINGULAR_G4|LINK]]
  |-
  |SUBMISSION
  |
* MIT - [https://mit.ilabsolutions.com/service_item/new/3381?spt_id=3863 ilabs]
  |-
|}
 
Minimum reads per lane are guaranteed if:
*the BMC has performed quality control,
*the samples are high-complexity, especially for the first 10 nt of read 1, and
*submitted libraries are at least 2 nM.'''
=== Workflow ===
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.
 
===Run Requirements===
<OL>
<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.
<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.
|
== The G4 Platform ==
{| class="wikitable" border=1
!width=100| SPEC
!width=250| G4
|-
|'''SEQUENCER'''||[[Image:2023_Singular_G4.jpg|center|200px]]
|-
| '''READS/LANE'''<BR> Low number is minimum per lane for standard libraries||
* F3 per Lane: 50-100 M
|-
|'''RUN TIME'''||
* 50 PE 11 hours
* 150 PE 24 hours
|-
|'''KITS AVAILABLE'''||
* 100nt
* 300nt
|-
|'''KNOWN APPLICATIONS'''||
* RNAseq including single cell
* Gene Expression
* Exome
* Target Enrichment
* Metagenomics
|-
|'''KEY NOTES'''||
* 4-color chemistry
* Patterned flow cell
* 4-lanes per flow cell
* 4-flow cells per run
* Index replacement or anchor expansion of Illumina libraries
|-
|'''THANKS TO'''||
* Singular Genomics
|}

Latest revision as of 00:52, 21 April 2026