- Barcoded PCR Primers for multiplexing contain embedded index sequence
- Up to 48 multiplexed samples
- 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® platforms
Note: these barcoded primers are provided in the NEXTflex Small RNA-Seq Kit v3 and do not need to be purchased separately.
Four sets of 12 NEXTflex™ Small RNA Barcode Primers, with embedded index sequences are available offering an improved multiplexing workflow and increased flexibility. This automation-friendly format enables multiplexing of up to 48 samples. The ability to pool samples in an efficient way significantly decreases hands on time while providing robust data quality. The primer barcoding system utilizes a 6 nt index to differentiate up to 48 different samples on a single flow cell lane.
48 barcoded primers are divided into 4 different kits (A-D) containing 12 unique barcodes for multiplexing. Each set of NEXTflex Small RNA Barcodes Primers contains 96 reactions worth of barcodes.
Selected Citations Referencing the NEXTflex Small RNA Barcode Primers:
Ahmed R, Chang Z, Younis A.E. et al. (2013) Conserved miRNAs Are Candidate Post-Transcriptional Regulators of Developmental Arrest in Free-Living and Parasitic Nematodes. Genome Biol Evol (2013) 5 (7): 1246-1260. doi: 10.1093/gbe/evt086.
Ennajdaoui, H., et al. (2016) IGF2BP3 Modulates the Interaction of Invasion-Associated Transcripts with RISC. Cell Reports. doi: 10.1016/j.celrep.2016.04.083.
Goodarzi, Hani, et al. (2014) Metastasis-suppressor transcript destabilization through TARBP2 binding of mRNA hairpins. Nature. doi:10.1038/nature13466.
Licciardello, G. et al. (2015) Deep sequencing and analysis of small RNAs in sweet orange grafted on sour orange infected with two citrus tristeza virus isolates prevalent in Sicily. Archives of Virology. doi: 10.1007/s00705-015-2516.
Maczuga, P. et al. (Jan 2013) Embedding siRNA sequences targeting Apolipoprotein B100 in shRNA and miRNA scaffolds results in differential processing and in vivo efficacy. Molecular Therapy (2013); 21 1, 217-227. doi: 10.1038/mt.2012.160
Nakashe, P. et al. (2011) Adapter Dimer Reduction in High-Throughput microRNA Profiling. OMICS 1(1): 6-11.
Prakash, P., Ghosliya, D. and Gupta, V. (2014) Identification of conserved and novel microRNAs in Catharanthus roseus by deep sequencing and computational prediction of their potential targets. Gene. doi. 10.1016/j.gene.2014.10.046.
Smith, O. et al. (2014) Genomic methylation patterns in archaeological barley show de-methylation as a time-dependent diagenetic process. Scientific Reports 4:5559 doi:10.1038/srep05559.
Velthut-Meikas A, Simm J, Tuuri T, Tapanainen JS, Metsis M, Salumets A (2013) Small RNA-seq of Human Granulosa Cells Reveals miRNAs in FSHR and Aromatase Genes. Mol. Endocrinol. 2013; 27:1128-1141.
Each of the NEXTflex™ Small RNA Barcode Primers sets contains 12 unique barcodes, enabling the user to multiplex up to 48 samples per flow cell lane. These kits ship on dry ice.
NEXTflex™ Barcode Primers
Required Materials Not Provided
total RNA or small RNA enriched from total RNA in nuclease-free water
100% Ethanol (stored at -20°C)
70% Ethanol (stored at room temperature)
3 M NaOAc, pH 5.2
2, 10, 20, 200 and 1000 µL pipettes
RNase-free pipette tips
Nuclease-free 1.5 mL microcentrifuge tubes
Thin wall nuclease-free 0.5 mL microcentrifuge tubes
Thin wall nuclease-free PCR tubes
15% TBE PAGE gel (1.0 mm)
1X TBE buffer
Clean razor or scalpel
Nucleic acid stain such as SYBR Gold® (Thermo Fisher Scientific®)
UV transilluminator or gel documentation instrument
Gel Electrophoresis apparatus
Electrophoresis power supply
RNA Clean® & Concentrator™ 5 Kit (Zymo Research® Cat # R1015) / RNeasy® MinElute® Cleanup (Qiagen® Cat # 74204)
0.45µm, 2 mL Spin-X® Centrifuge tube (Sigma® Cat # CLS8162)