Page Comparison

Input amounts

1ng - 500ng*

*low input protocol modifications available for 1 ng - 10 ng.

10ng - 120ng per sample into First Strand Synthesis.

80ng - 960ng per pool into Second Strand Synthesis.

RNA quality

High and low quality RNA is acceptable, as well as FFPE RNA.

Medium to high quality RNA required (i.e., RIN >6).

Blood samples

Compatible with Globin Block Modules
(Cat. No. 070 (Homo sapiens) and 071 (Sus scrofa)).

Not recommended.

Unique Molecular Identifiers (UMIs)

Optional with UMI Second Strand Synthesis Module (Cat. No. 081).
UMIs are 6 nt long (plus a 4 nt spacer) and added during Second Strand Synthesis.

Included.

UMIs are 10 nt long and added during First Strand Synthesis.

First Strand Synthesis Primer

Oligo(dT) primer with partial Illumina adapter sequence.

Oligo(dT) primer with 12 nt i1 barcode, 10 nt UMI, and partial Illumina adapter sequence.

Library Quality Control (QC)

QC individual libraries and pool (on sequencer flow cell) at equimolar ratio.

QC pooled library; individual library QC not possible.

Average Library Size

~335 – 456 bp (based on UHRR)*

*Can vary based on RNA quality and sample type.

~650 bp (based on UHRR)*

*Can vary based on RNA quality and sample type.

Sequencing mode

Single Read (SR)

Paired End Read (PE)
Read 1: insert
Read 2: UMI and i1 barcode

Demultiplexing

i5/i7 demultiplexing required.

If only using Lexogen 12 nt UDI in your sequencing lane (e.g., with other Lexogen libraries), we recommend iDemux for demultiplexing and optimal error correction.

i5/i7 demultiplexing optional (if UDI are used).

i1 demultiplexing required (iDemux recommended).

If only using Lexogen 12 nt UDI in your sequencing lane (e.g., with other Lexogen libraries), we recommend iDemux for demultiplexing and optimal error correction.