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Lexogen 12 nt UDI Sets -
Tag your libraries with ultimate accuracy

Lexogen 12 nt Unique Dual Index (UDI) System for RNA-Seq

Lexogen’s 12 nt Unique Dual Indices (UDIs) are introduced at the PCR step and enable unique barcoding of up to 384 samples per lane run. Lexogen’s 12 nt UDIs feature superior error correction for maximal sequencing data output.

Lexogen’s 12 nt UDIs are available as convenient bundles or Add-on sets, compatible with the following Lexogen library prep solutions:

Performance

Lexogen’s 12 nt UDIs enable maximized NGS data output due to sophisticated index design, superior error correction, and adjustable read-out length accommodation for each experimental scale.
Multiplexed RNA-Seq can suffer from read misassignments due to Index Hopping and random Index Sequence Errors, potentially compromising data accuracy. Using Unique Dual Indexes (UDI), like Lexogen’s 12 nt UDI, effectively resolves these issues by uniquely identifying and allowing for the removal of misassigned reads, and enabling robust error correction to maximize data output (Figure 1.).
Figure 1 | The effects of Index Hopping and Index Sequence Errors in a pool of libraries with Unique Dual Indexing. Read mis-assignment caused by Index Hopping can be avoided by using Unique Dual Indexing (UDI). Reads with hopped indices are irreversibly discarded (C). Reads with random Index Sequence Errors resulting in an index not present in the pool are classified undetermined. Accurate error correction can rescue most of these reads making them available for downstream data analysis (B). The percentage values were derived from an RNA-Seq experiment pooling 96 libraries with Lexogen’s 12 nt UDIs and full 12 nucleotide index read-out on an Illumina NextSeq500.

The Lexogen UDI 12 nt Unique Dual Indices are 12 nucleotides (nt) long and designed to maximize inter-index distance for different sample numbers and index read-out lengths. In a typical experiment using the full 12 nt index read-out, around 9.1 % of the initial raw reads contain a random Index Sequence Error (Fig. 2A). This renders them undetermined, hence removing these reads from downstream analysis.

Lexogen’s advanced index design enables the rescue of 76 % of these undetermined reads increasing the useful output thereby increases to 97.8 % of the initial reads, an unprecedented performance due to the cutting-edge index design (Fig. 2B).

Figure 2 | Maximizing read output with Lexogen’s 12 nt UDIs and error correction. 96 multiplexed libraries were sequenced on an Illumina NextSeq500 with 12 nt UDI read-out. A) In a standard RNA-Seq experiment a significant number of reads is undetermined (orange) due to random Index Sequence Errors. B) Lexogen’s 12 nt Unique Dual Indices are optimized for maximal error correction with highest accuracy. Lexogen’s Error Correction Tool allows almost 7% of originally undetermined reads to be confidently rescued and correctly assigned to the respective library.

Lexogen’s 12 nt Unique Dual Indexes (UDIs) offer flexible read-out lengths of 8, 10, or 12 nucleotides for unique barcoding and to accommodate diverse multiplexing needs, ensuring accuracy and reliable error correction based on experiment type, sequencing equipment, desired read depth, and / or the number of pooled libraries. Longer 12 nt UDIs, while requiring slightly more sequencing cycles, provide enhanced error correction capabilities for more complex index sequence errors, eliminating the need for multiple indexing sets.

Lexogen’s 12 nt Unique Dual Indexes (UDIs) are designed with a nested approach, optimizing error correction and inter-index distance for various multiplexing scales, from small to extreme. All UDI subsets are nucleotide-balanced for superior cluster identification in NGS runs.
Figure 3 | Distance and error correction in Lexogen’s nested 12 nt UDI sets. Illustration of inter-index distance (D) and number of possible error corrections (ec) in a nested index set with 12 nt read-out. An optimized set of 384 indices contains a subset of 96 indices with larger distances and enhanced error correction. Within these 96 indices, a subset of 24 indices is optimized even further, while a 4 index subset features the highest possible inter-index distance and error correction capacity.
The Lexogen UDI 12 nt Unique Dual Index system adapts to the user’s needs while always providing the highest inter-index distance and maximal error correction capacity. Read mis-assignment due to Index Hopping is avoided, and Index Sequence Errors can be corrected with highest accuracy. Thereby, the system provides the optimized indexing solution for current and future barcoding requirements.
Subset12 nt10 nt8 nt
DecDecDec
384413121*
     └ 96524131
           └ 24625241
                 └ 4736252

Table 1 | Comparison of distance and error correction capacity in Lexogen’s nested 12 nt UDI sets with 8, 10, and 12 nt read-out. Inter-index distance (D) and number of errors that can be corrected (ec) are compared for subsets of 384, 96, 24, and 4 libraries and the three possible read-out lengths. For smaller subsets (up to 96 samples) a read-out of 8 or 10 nt allows correction of one error and thus recovery of additional reads. Larger subsets require a read-out of 10 or 12 nt to benefit from the error correction. The ec values represent the number of all errors (including substitutions, insertions, and deletions) that can be confidently corrected, except for *. In this case error correction can only address substitutions.

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Lexogen 12 nt Unique Dual Index System (UDI) for RNA-Seq

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Ordering Information

Lexogen’s 12 nt UDIs are available as UDI Sets A1 – A4 and UDI Set B1, as part of convenient bundles with QuantSeq, CORALL, or LUTHOR HD library prep kits, or as standalone Add-on kits.

QuantSeq FWD - Catalog Numbers
Cat. No. Product Name
QuantSeq FWD
191.96QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Set A1, (UDI12A_0001-0096), 1 rxn/UDI
192.24QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Set B1, (UDI12B_0001-0024), 1 rxn/UDI
192.96QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Set B1, (UDI12B_0001-0096), 1 rxn/UDI
193.384QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Sets A1-A4, (UDI12A_0001-0384), 1 rxn/UDI
194.96QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Set A2, (UDI12A_0097-0192), 1 rxn/UDI
195.96QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Set A3, (UDI12A_0193-0288), 1 rxn/UDI
196.96QuantSeq 3’ mRNA-Seq V2 Library Prep Kit FWD with UDI 12 nt Set A4, (UDI12A_0289-0384), 1 rxn/UDI
Cat. No. Product Name
CORALL mRNA-Seq
177.96CORALL mRNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, (UDI12A_0001-0096), 96 preps
178.96CORALL mRNA-Seq V2  Library Prep Kit with UDI 12 nt Set A2, (UDI12A_0097-0192), 96 preps
179.96CORALL mRNA-Seq V2 Library Prep Kit with UDI 12 nt Set A3, (UDI12A_0193-0288), 96 preps
180.96CORALL mRNA-Seq V2 Library Prep Kit with UDI 12 nt Set A4, (UDI12A_0289-0384), 96 preps
181.96CORALL mRNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, (UDI12B_0001-0096), 96 preps
182.384CORALL mRNA-Seq V2 Library Prep Kit with UDI 12 nt Sets A1-A4 (UDI12A_0001-0384), 384 preps
Cat. No. Product Name
CORALL RNA-Seq Stand-alone Kits without rRNA Depletion, including Lexogen 12 nt UDI Sets
171.24CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, (UDI12A_0001-0024), 24 preps
171.96CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, (UDI12A_0001-0096), 96 preps
172.96CORALL RNA-Seq V2  Library Prep Kit with UDI 12 nt Set A2, (UDI12A_0097-0192), 96 preps
173.96CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A3, (UDI12A_0193-0288), 96 preps
174.96CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A4, (UDI12A_0289-0384), 96 preps
175.24CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, (UDI12B_0001-0024), 24 preps
175.96CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, (UDI12B_0001-0096), 96 preps
176.384CORALL RNA-Seq V2 Library Prep Kit with UDI 12 nt Sets A1-A4 (UDI12A_0001-0384), 384 preps
Cat. No. Product Name
CORALL RNA-Seq with rRNA Depletion, including Lexogen 12 nt UDI Sets
183.24RiboCop (HMR) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, (UDI12A_0001-0024), 24 preps
183.96RiboCop (HMR) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, (UDI12A_0001-0096), 96 preps
184.24RiboCop (HMR) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, (UDI12B_0001-0024), 24 preps
184.96RiboCop (HMR) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, (UDI12B_0001-0096), 96 preps
Cat. No. Product Name
LUTHOR HD
221.24LUTHOR High-Definition Single-Cell 3’ mRNA-Seq Kit with UDI 12 nt Set B1, (UDI12B_0001-0024), 24 preps
221.96LUTHOR High-Definition Single-Cell 3’ mRNA-Seq Kit with UDI 12 nt Set B1, (UDI12B_0001-0096), 96 preps
Cat. No. Product Name
UDI Add-on Kits (UDI plates only), e.g., for QuantSeq-Pool and LUTHOR HD Pool
101.96Lexogen UDI 12 nt Set A1 (UDI12A_0001-0096), 1 rxn/UDI
102.96Lexogen UDI 12 nt Set A2 (UDI12A_0097-0192), 1 rxn/UDI
103.96Lexogen UDI 12 nt Set A3 (UDI12A_0193-0288), 1 rxn/UDI
104.96Lexogen UDI 12 nt Set A4 (UDI12A_0289-0384), 1 rxn/UDI
105.96Lexogen UDI 12 nt Set B1 (UDI12B_0001-0096), 1 rxn/UDI
156.384Lexogen UDI 12 nt Sets A1-A4 (UDI12A_0001-0384), 1 rxn/UDI
Cat. No. Product Name
UDI Add-on Kits (UDI plate and library amplification module)
198.96Lexogen UDI 12 nt Unique Dual Indexing V2 Add-on Kit, Set A1 (UDI12A_0001-0096), 1 rxn/UDI
199.96Lexogen UDI 12 nt Unique Dual Indexing V2 Add-on Kit, Set A2 (UDI12A_0097-0192), 1 rxn/UDI
200.96Lexogen UDI 12 nt Unique Dual Indexing V2 Add-on Kit, Set A3 (UDI12A_0193-0288), 1 rxn/UDI
201.96Lexogen UDI 12 nt Unique Dual Indexing V2 Add-on Kit, Set A4 (UDI12A_0289-0384), 1 rxn/UDI
202.96Lexogen UDI 12 nt Unique Dual Indexing V2 Add-on Kit, Set B1 (UDI12B_0001-0096), 1 rxn/UDI
203.384Lexogen UDI 12 nt Unique Dual Indexing V2 Add-on Kit, Sets A1-A4 (UDI12A_0001-0384), 1 rxn/UDI

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