The Libraries prepared with the QuantSeq 3' mRNA-Seq REV kit is appropriate for require a Custom Sequencing Primer Version 5 (CSPV5), for Read 1 sequencing. Therefore, REV libraries can be sequenced on platforms that support the use of custom sequencing primers for Read 1.
These platforms include: NextSeq 500 / 550, NextSeq 1000 / 2000, HiSeq 2000 / 2500 / 3000 / 4000, HiSeq 3000, HiSeq 4000, GAIIX, and MiSeqWhat about iSeq and NovaSeq?Also NextSeq 2000. We do not recommend to sequence QuantSeq REV libraries on NextSeq 500 or NextSeq 550 Illumina platforms. The polyadenylation signal located at the 3' end, which is quite similar between transcripts, will result in a reduced complexity at the beginning of the reads leading to a lower sequencing qualityNovaSeq 6000, MiSeq, and AVITI instruments.
QuantSeq REV libraries are not compatible with the iSeq 100 instrument, as this platform does not support the use of custom sequencing primers.
ATTENTION! Always consult the Illumina Custom Sequencing Primer guide or the AVITI System Workflow Guide for the relevant instrument.
NOTE: Lexogen does not have direct experience with all of these platforms and reagent versions. These recommendations are based on custom sequencing primer compatibility for these systems. Please inquire at support@lexogen.com for updates, or contact Illumina technical support / Element technical support for further details of custom primer use for your specific sequencing setup.
Please also check loading amount guidelines for QuantSeq REV libraries here Do you have recommended loading concentrations for Lexogen libraries on Illumina sequencing instruments? For loading on Illumina instruments, in general, we recommend loading QuantSeq REV libraries at the higher range of the Illumina-provided specifications.