BioMicroCenter:Element Sequencing

Element Sequencing

The Center currently hosts an AVITI24 platform from Element Biosciences. AVITI24 platform supports both sequencing and cytoprofiling. For most sequencing needs, the Center will use Element's Freestyle chemistry kits.

Service	Element Sequencing
SUBMISSION	Illumina OR Element libraries
MIN VOLUME	15 uL
MIN CONCENTRATION	2 nM
INCLUDED SERVICES	Quality Control:Fragment Analyzer and qPCR Element Sequencing Demultiplexing
ADDITIONAL SERVICES	Quality Control Element library preperation
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

Illumina library is preferred. Element libraries require a 5'-phosphate on the forward end in order to circularize and sequence. Compared to an Illumina platform, AVITI24 reads are similar in read direction to Illumina's Forward Strand workflow for Dual Indexed Libraries.

Minimum reads per lane are guaranteed if:

• the BMC has performed quality control,
• the samples have high diversity, especially within the first 5 bases of read 1, and
• submitted libraries are at least 2 nM.

Lane-by-lane Notes

• The wait is variable
• Custom primers must be compatible with TruSeq, Nextera, TruSeq, smRNA and Element sequencing primers

All other requests require a full flowcell. 300PE kits only have a single lane.

• Min custom primer submission: 70 uL at 100 uM for Read 1, all other reads require 40 uL at 100 uM.

The AVITI24 Platform

	AVITI24	NOTES
SEQUENCER
READS/FLOWCELL Low number is minimum per lane for standard Illumina libraries.	300M HQ read pairs PF/flowcell
RUN TIME	75 PE 24 hours 150 PE 38 hours 300 PE 60 hours
KITS AVAILABLE	2x75 Cloudbreak FS High 2x150 Cloudbreak FS High 2x300 Cloudbreak FS High	- Single end supported - each kit comes with 34 additional cycles
KNOWN APPLICATIONS	Whole Genome Sequencing RNA Sequencing 16S Single-cell Target Sequencing and Enrichment Metagenomics
KEY NOTES	4-color chemistry Random flow cell 2-lanes per flow cell Q40+ > 80% Low-binding surface chemistry Adept Workflow for adaptation of Illumina libraries
THANKS TO	Element Biosciences

Aviti24 circularizes linear Element or Illumina libraries onboard, and then performs rolling circle amplification to generate thousands of copies of the original template on a single strand. The strand is then packaged up into a polony. These polonies are washed with a complex of primers, polymerase, and dye-labelled avidites. Avidites are multivalent-nucleotide-ligands on dye labeled cores capable of binding to multiple sites to form a polymerase–polymer–nucleotide complex within a polony. After an imaging step, the avidites are washed away and the growing read strand incorporates a single nucleotide that is reversibly blocked on the 3' end. The polonies are again loaded with avidites, repeating the process over many cycles.

The AVITI24 can run 2 flow cells per run, each hosting 2 independently-addressable lanes. Both lanes are washed with the same reagents. Element currently offers kits with 150 and 300 cycles, both supporting 2 lanes, as well as a 600 cycle kit that supports a single lane. Index hopping rates are virtually nil, limiting the need for UDI's. The AVITI24 can also perform direct in sample sequencing which includes spatial imaging of RNA, proteins, phospho-proteins, and cell morphology using Teton chemistry for cytoprofiling.