BioMicroCenter:Singular Sequencing: Difference between revisions

Revision as of 13:57, 28 May 2025

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

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.

Run Requirements

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.

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

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

@@ Line 44: / Line 44: @@
 *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.'''
+*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===