Below you can find a table of advised loading concentrations for different Lexogen library pools on common Illumina sequencing platforms. If specified ranges are provided these are based either on Lexogen’s own validation of loading concentrations, and/or feedback provided by Lexogen customers, and are known to contribute to optimal sequencing quality and output. These recommendations apply to pure lanemixes per library type. If you are running lanemixes featuring a mixture of different library prep types, loading concentrations may need to be optimized specifically.
PhiX spike-in recommendations are also provided. Please note PhiX should not be added to QuantSeq REV runs. Due to the use of a custom sequencing primer (CSP) PhiX will not be detected.
NOTE: Where no specific loading concentration / PhiX details are provided please refer to the Illumina loading concentration recommendations for the specific instrument and/or flow cell. We can generally recommend starting at the higher end of these Illumina loading concentration ranges.
QuantSeq FWD (Cat. No.s 191-196) & QuantSeq-Flex (Cat. No.s 166, 028) | QuantSeq FWD with UMIs (Cat. No.s 191-196 + 081) | QuantSeq-Pool (Cat. No. 139) | QuantSeq REV* (Cat. No. 225) | CORALL (Cat.no.s 171-176, 177-182, 183-186) | LUTHOR HD (Cat. No.s 204, 221) | Small RNA-Seq (Cat. No.s 052, 058) | |
|---|---|---|---|---|---|---|---|
NextSeq 500 / 550 2 | 2 - 2.5 pM4 (+ 1 - 5% PhiX) | 1.8 - 2 pM4 (+ 15 - 30% PhiX) 1 | 2 - 2.5 pM4 (+ 1 - 5% PhiX) | 2 - 3 pM4 | 2.2 - 2.7 pM4 (+ 1 - 5% PhiX) | 2 - 2.5 pM3 (+ 1 - 5% PhiX) | 2.6 pM4 (5 % PhiX) |
NextSeq 2000 3 | 650 - 750 pM (+ 1 - 5% PhiX) | 750 pM (+ 30% PhiX) 1 | 700 - 800 pM (+ 1 - 5% PhiX) | ** | 650 - 750 pM (+ 1 - 5% PhiX) | ** | 650 pM |
MiSeq | 6 – 15 pM (+ 1 - 5% PhiX) | 11 pM (+ 5% PhiX) 1 | ** | 6 – 15 pM | ** | 6 - 15 pM (+ 1 - 5% PhiX) | 15 – 20 pM |
MiniSeq | 1.3 – 1.8 pM (+ 1 - 5% PhiX) | ** (+ 15 - 30% PhiX) 1 | ** | ** | ** | 1.3 - 1.8 pM (+ 1 - 5% PhiX) | ** |
NovaSeq 6000, X, X Plus | 200 - 400 pM (+ 1 - 5% PhiX) | ** (+ 15 - 30% PhiX) 1 | ** | ** | ** | Standard: 300 - 500 pM, | ** |
iSeq 100 | ** | ** (+ 15 - 30% PhiX) 1 | ** | Not compatible!5 | ** | ** | ** |
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