Unique dual index adapters for multiplexing up to 192 Illumina® sequencing libraries
- Mitigate index hopping or spread of signal that can occur during sequencing
- Each lot of the NEXTFLEX® Unique Dual Index Barcodes is functionally validated and tested for index purity by sequencing
- Compatible with both paired-end and single-read Illumina® sequencing
For research use only. Not for use in diagnostic procedures.
The NEXTFLEX® Unique Dual Index Barcodes are barcoded adapters, for sequencing on Illumina® platforms, that provide unprecedented data security in sequencing applications. Multiplexing with NEXTFLEX® Unique Dual Index Barcodes drastically increases scale while reducing costs by allowing the user to pool multiple libraries in a single flow cell lane. These barcoded adapters can be used with single read or paired-end read sequencing, and are compatible with NEXTFLEX® library prep kits designed for Illumina® sequencing, or other workflows for Illumina® sequencing that involve the ligation of adapters to adenylated fragments.
Unique dual indexes prevent data ambiguity caused by index hopping
The NEXTFLEX® Unique Dual Index Barcodes utilize two unique 8 nt indexes on each UDI adapter. The 8 nt indices are designed with a Hamming Distance of at least three throughout the entire series, permitting error correction. Each NEXTFLEX® Unique Dual Index Barcode comprises the entire flow cell binding site, sequencing primer binding site, and index sequences, making PCR enrichment optional. Patterned flow cells, such as those found on the NovaSeq® and HiSeq® 3000/4000 platforms, have been recognized to suffer from increased rates of sample mis-assignment during sequencing. The use of adapters with unique dual indices during sequencing prevents such mis-assigned reads from appearing in final data sets, allowing for the highest assurance of data integrity.
Reduce run-to-run sample carryover and sample crosstalk with UDIs
Even users who don’t routinely perform high-throughput multiplexing benefit from the superior design of the NEXTFLEX® Unique Dual Index Barcodes. The ability to avoid using the same set of indices consecutively reduces concerns about run-to-run sample carryover and sample crosstalk. The NEXTFLEX® Unique Dual Index Barcodes are designed to specifically address the index hopping, or spread of signal, phenomenon associated with Illumina® platforms that utilize a patterned flow cell.
Each NEXTFLEX® Unique Dual Index Barcodes Kit contains 96 unique dual-index barcodes, enabling users to multiplex up to 96 samples per flow cell lane. Each lot of the NEXTFLEX® Unique Dual Index Barcodes is functionally validated and tested for index purity by sequencing. NEXTFLEX® Unique Dual Index Barcodes are supplied in 96-well plates with each well containing 2 reactions of each barcode. These kits ship on dry ice.
NEXTFLEX® Unique Dual Index Barcodes (25 µM)
NEXTFLEX® Primer Mix (12.5 µM)
Q: Are the NEXTflex® unique dual index barcodes sequence-verified for purity?
A: Yes, each lot of the NEXTflex® unique dual index barcodes undergoes stringent QC procedures that include verification of the index sequence purity by sequencing on a MiSeq® instrument.
Q: Are the NEXTflex® unique dual index barcodes functionally validated?
A: Yes, each lot of the NEXTflex® unique dual index barcodes undergoes stringent QC procedures that include functional validation of the barcoded adapter by making a library.
Q: What is the lead time when I order the NEXTflex® unique dual index barcodes?
A: The NEXTflex® unique dual-index barcodes are in-stock and ready-to-ship.
Q: Why are the NEXTflex® unique dual index barcodes only supplied at two reactions of each barcode in the 96-well plate?
A: The NEXTflex® unique dual index barcodes are supplied in aliquots of two reactions of each barcode to reduce the potential for contamination, which increases with repeated use of the same plate.
Q: Must I perform paired-end sequencing with NEXTflex® Unique Dual Index Barcodes?
A: No, the NEXTflex® Unique Dual Index Barcodes are compatible with paired-end and single-read sequencing runs. The I7 and I5 indexes are read in separate sequencing events than Read 1 and Read 2.
Q: Can I use the NEXTflex® Unique Dual Index Barcodes in single index experiments?
A: Yes, you can configure your sequencing run to only read the I7 index when you set up the run.
Sequences of NEXTflex Dual-Indexed DNA Barcode Indexes
Instructions for installing NEXTflex Barcode Indices in Illumina Experiment Manager
NEXTflex Unique Dual Index Barcodes Manual