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QuantSeq-FWD | QuantSeq-Pool | |
---|---|---|
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 | Not recommended. |
Unique Molecular Identifiers (UMIs) | Optional with UMI Second Strand Synthesis Module (Cat. No. 081). | Included. |
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 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) |
Demultiplexing | i5/i7 demultiplexing required.
| i5/i7 demultiplexing optional. i1 demultiplexing required (iDemux recommended). If only using Lexogen 12 nt UDI in your sequencing lane, we recommend iDemux for demultiplexing and optimal error correction. |
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