INDEXED RNA-SEQ ADAPTERS FOR MULTIPLEXING LIBRARIES ON ILLUMINA® PLATFORMS
NEXTFLEX-96™ RNA-Seq Barcodes
- Index contained within adapter sequence eliminates the need to perform PCR to add flow cell binding sequences
- Multiplex up to 96 libraries
- Considerably reduce your per-sample sequencing cost by barcoded multiplexing
- Increase your sequencing scale by pooling 100s of samples on a single flow cell
- Compatible with Illumina® next-generation sequencing platforms
- Compatible with the NEXTFLEX® Rapid RNA-Seq Kit and the NEXTFLEX® Rapid Directional RNA-Seq Kit
- Not compatible with the NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0
(in 96-well plate)
For research use only. Not for use in diagnostic procedures.
Increase Scale while Reducing Cost
The NEXTFLEX-96™ RNA-Seq Barcodes are adapters that were designed for use with the NEXTFLEX® Rapid RNA-Seq Kit and the NEXTFLEX® Rapid Directional RNA-Seq Kit. These barcodes are not recommended for use with the new NEXTFLEX® Rapid Directional RNA-seq kit 2.0.
The NEXTFLEX-96™ RNA-Seq Barcodes can be used to provide flexibility and high-throughput capabilities in sequencing applications. They significantly increase scale while reducing costs by allowing the user to pool multiple library preparations in a single flow cell lane. The NEXTFLEX-96™ RNA-Seq Barcodes kits accomplish this by using an indexed adapter with an 8 nt unique sequence. This allows for proper differentiation between samples, preventing poor reads from single base errors introduced during PCR. The NEXTFLEX® index is contained within the adapter sequence eliminating the need to perform PCR to add flow cell binding sequences. The NEXTFLEX-96™ RNA-Seq Barcode index sequences are designed to have a hamming distance of 4, allowing discrimination of different adapters even in the case of an error in the barcode read. These adapters are supplied a 96-well format.
Do you want to index you samples but don’t want to use 96 different barcodes? We also offer the NEXTFLEX® RNA-Seq Barcodes in sets of 6, 12, 24 and 48 barcodes.
For larger volume requirements, customized and bulk packaging is available. Please contact [email protected] for further information.
Avoiding Registration Failure with Low Level Multiplexing
Registration failure could occur if the color balance was not maintained between the red and green lasers (used to sequence A/C bases and G/T bases, respectively). Read our blog post, Tech Tips – Barcode Recommendations for Low Level Multiplexing, to learn how to avoid registration failure on an Illumina® sequencer caused by lack sufficient index sequence diversity.
- NEXTFLEX® RNA-Seq Barcode Adapter (0.6 µM)
- NEXTFLEX® Primer Mix (12.5 µM)
Selected Publications that Reference using the NEXTFLEX-96™ RNA-Seq Barcodes
- Harvey, C. T. et al. (2015) QuASAR: quantitative allele-specific analysis of reads. Bioinformatics. Doi: 10.1093/bioinformatics/btu802
- Moyerbrailean, G. A., et al. (2015) A high-throughput RNA-seq approach to profile transcriptional responses. Scientific Reports. 5: 14976. doi: 10.1038/srep14976.
- Takashima, Y. (2014) Resetting Transcription Factor Control Circuitry toward Ground-State Pluripotency in Human. Cell. 158: 6, 1254–1269.
The NEXTFLEX-96™ RNA-Seq Barcodes Kits contain 96 unique adapters compatible with Illumina® platforms, enabling the user to multiplex up to 96 RNA-seq samples. These kits ship on dry ice.
These barcodes are not recommended for use with the new NEXTFLEX® Rapid Directional RNA-seq kit 2.0. For early access evaluations of this kit, 8 reactions of barcodes will be provided at no cost.