BioMicroCenter:Singular Sequencing

Singular Sequencing

The Center currently hosts a G4 platform by 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.

Service	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	FASTQC Basic run metrics (alignment rate, complexity) Basic RNAseq metrics (where applicable) Basic paired end metrics (where applicable) Contamination checks
PRICING	LINK
SUBMISSION	MIT - ilabs

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.

Lane-by-lane Requirements

• Must be 50 or 150 paired-end dual indexed
• Pool must be compatible with TruSeq, Nextera, TruSeq, smRNA and Solexa sequencing primers

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.

All other requests, including use of custom sequencing primers require a full flowcell. Illumina libraries which have been converted in the Center will also require a full flowcell.

• Min custom primer submission: 20 uL at 100 uM

Index hopping for a standard set of Singular libraries

Percent Perfect Plot for Singular libraries 150PE

Index hopping on Singular runs has been low. The performance of an F3 flowcell mimics other short-read 4 color chemistry sequencers with patterned flowcells.

The G4 Platform

SPEC	G4
SEQUENCER
READS/LANE 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