AVITI Loading Guidelines
The loading guidelines provided below are for Lexogen libraries prepared using the Adept workflows for sequencing on the Element Bioscience AVITI sequencing platform. These guidelines are based on the current experience of Lexogen and Lexogen customers.
If loading concentrations are not provided for the size range, or Adept workflow used to prepare your libraries/lanemix, please consult the relevant AVITI System Workflow guide (according to the type of sequencing kit used and current OS version of your AVITI instrument).
If you have further questions or would like specific loading advice please contact support@lexogen.com and provide a copy of the linear lanemix size distribution (trace file, pre-circularization) and the type and percentage of each library included in the lanemix.
Library Type | Recommended Read Format (Seq. Kit)* | Custom Primers Required? | Average Size Range | CloudBreak Circular Adept v1.1 Libraries | CloudBreak Linear Adept Rapid PCR-Plus Libraries |
|---|---|---|---|---|---|
CORALL RTM | SR150; PE150 (2x75; 2x150) | No | 300 - 450 bp | 6 - 6.5 pM2 2 - 5% PhiX | |
CORALL RTL | PE-150 (2x150) | No | 500 - 600 bp | 8 - 8.5 pM2 2 - 5% PhiX | |
QuantSeq FWD | SR150 (2x75)1 | No | 250 - 400 bp | 5 - 6 pM4 2% PhiX | 12 pM4 |
400 - 500 bp | 7 - 8 pM2,4 2% PhiX | ||||
QuantSeq FWD-UMI | SR150 (2x75)1 | No | 250 - 400 bp | 7.5 - 8.5 pM4 2% PhiX for runs with <= 20% of UMI libraries in total lanemix. -for advice on PhiX for runs that contain >20% QuantSeq FWD-UMI libraries please contact support@lexogen.com | |
QuantSeq-Pool | PE125/25 (2x75) | No | 400 - 550 bp | 8.5 - 10 pM4 2% PhiX | |
Luthor HD | PE125/25 (2x75) | No | 500 - 600 bp | 8.5 - 10 pM4 2% PhiX | |
Mixed Lexogen libraries (excluding small RNA-Seq) | SR150; PE75; PE150 (2x75, 2x150) | No | 300 - 400 bp | 6 pM4 2% PhiX | |
SR150; PE75; PE150 (2x75, 2x150) | No | 500 - 700 bp | 9 - 11 pM4 2% PhiX | ||
Lexogen small RNA-Seq | SR75; PE75 (2x75) | Yes [R1, R2, I2] | 143 - 200 bp | Please contact support@lexogen.com for updated recommendations if you wish to sequence these library types on the AVITI. | |
QuantSeq REV 3' mRNA Seq | SR150 (2x75)1 | No - if combined with other Lexogen libraries at <= 10% of lanemix.4 | 250 - 400 bp | ||
*The 2 x 300 cycle kits are not recommended for sequencing Lexogen library types.
1 QuantSeq FWD libraries can also be sequenced in Paired End (PE) mode if required. Note this will cause a quality drop for read 2 which may lead to a lower % Pass Rilter (%PF) rate. In such cases, only Read 1 should be used for downstream QuantSeq data analysis.
2 Loading concentrations evaluated directly by Lexogen for stand-alone runs using only this library type.
3 Loading concentrations evaluated by Lexogen customers/Element Biosciences - stand-alone runs using only this library type.
4 Suggested loading only based on library size using suggested Element Bioscience loading range, or from sequencing of library type included in mixed library lanemix.