- Explore our NEW! NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0 for an updated chemistry that delivers high performance.
- 2 hours faster than traditional Illumina® stranded RNA-Seq library prep protocols
- Provides precise measurement of strand orientation (>99%)
- >10 ng – 1 µg total RNA for enrichment by NEXTFLEX® Poly(A) Beads 2.0 or ~ 1 ng – 100 ng isolated mRNA or rRNA-depleted RNA
- Minimal hands-on time required
- Streamlined protocol reduces sample loss
- Utilizes dUTP-based methodology
- Robust, reliable performance
- Up to 96 barcodes available for multiplexing
- Automation protocols are available for the Sciclone® NGS and NGSx workstations
- Functionally validated with Illumina® sequencing platforms
- Allows for single read or paired end transcriptome sequencing
Improve Transcript Annotation & Improve Read Alignment
The NEXTFLEX® Rapid Directional RNA-Seq Kit provides an easy and flexible solution for generating single read, paired-end and multiplexed stranded libraries from mRNA or rRNA depleted total RNA. This kit can be used to obtain “stranded” information which identifies the specific DNA strands a given RNA transcript was derived from with >99% strand specificity. This stranded information improves transcript annotation and improves read alignment, reducing per sample sequencing costs. dUTP-based strand orientation also enables the detection of antisense expression regulatory relationships and allows for the discovery of novel ncRNA.
The NEW! NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0 now available!
Cost Effective, Stranded RNA Library Prep Kit Bundles
To simplify your mRNA sequencing, PerkinElmer now offers affordable bundles containing the NEXTFLEX® Rapid Directional RNA-Seq library prep reagents, NEXTFLEX® RNA-Seq Barcodes and NEXTFLEX® Poly(A) Beads. There are two different versions of this kit. They each contain the 48 rxn NEXTFLEX® Rapid Directional RNA-Seq Kit, NEXTFLEX® Poly(A) Beads (48 rxns) and 24 unique NEXTFLEX® RNA-Seq Barcodes (in aliquots of 2 reactions each).
Improved Rapid Stranded RNA-Seq Offers Higher Library Yields and Better Mapped Reads
The whitepaper, Improved Rapid Stranded RNA-Seq Offers Higher Library Yields and Better Mapped Reads, illustrates a detailed comparison of the NEXTFLEX® Rapid Directional RNA-Seq Kit with a competitor’s fast directional library prep kit. The NEXTFLEX® Rapid Directional RNA-Seq Kit excelled at rRNA removal, library yield and overall quality metrics.
Robust Library Prep with Low Input Amounts of Total RNA
Our whitepaper, Constructing High Quality RNA-Seq Libraries from Limited Amounts of Total RNA, illustrates the robustness of the NEXTFLEX® Rapid Directional RNA-Seq Kit used along with NEXTFLEX® Poly(A) Beads for mRNA isolation and library preparation from input as low as 10 ng of total RNA.
Automation Protocols for Stranded RNA Library Prep
Optimized automation protocols for stranded RNA library prep using the NEXTFLEX® Rapid Directional RNA-Seq Library Preparation Kits are available for the Sciclone® NGS and NGSx Workstations. Download the Sciclone NGS and NGSx Workstation Automation Guides for the NEXTFLEX® Rapid Directional RNA-Seq Kit.
Flexible Multiplexing Options for Illumina® RNA Sequencing
The NEXTFLEX® Rapid Directional RNA-Seq Library Prep Kit was designed to be used in conjunction with the NEXTFLEX® RNA-Seq Barcodes or NEXTFLEX-96™ RNA-Seq Barcodes for multiplexing. These single index barcodes are available in sets of 6, 12, 24, 48 and 96 barcodes.
Poly(A) Beads for mRNA Purification
The NEXTFLEX® Poly(A) Beads 2.0 offer a convenient method, validated with the NEXTFLEX® Rapid Directional RNA-Seq Kit, for purification of pure, intact mRNA.
- NEXTFLEX® RNA Fragmentation Buffer
- NEXTFLEX® First Strand Synthesis Primer
- NEXTFLEX® Directional First Strand Synthesis Buffer Mix
- NEXTFLEX® Rapid Reverse Transcriptase
- NEXTFLEX® Directional Second Strand Synthesis Mix
- NEXTFLEX® Adenylation Mix
- NEXTFLEX® Ligation Mix
- NEXTFLEX® RNA-Seq Barcode Adapter 1 (0.6 µM)
- NEXTFLEX® Uracil DNA Glycosylase
- NEXTFLEX® Primer Mix (12.5 µM)
- NEXTFLEX® PCR Master Mix
- Nuclease-free Water
- Resuspension Buffer
REQUIRED MATERIALS NOT PROVIDED
- 10 ng – 1 µg total RNA for enrichment by NEXTFLEX® Poly(A) Beads 2.0 or ~ 1 ng – 100 ng isolated mRNA or rRNA-depleted RNA
- DynaMag™-2 Magnet (Life Technologies Cat # 123-21D)
- Commercial kits for rRNA depletion (optional, if not using poly(A) enrichment)
- 100% Ethanol (stored at room temperature)
- 80% Ethanol ( freshly prepared )
- 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
- 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) / or / similar
- Adhesive PCR Plate Seal (BioRad, Cat # MSB1001)
- Agencourt AMPure XP 60 mL (Beckman Coulter Genomics, Cat # A63881)
- Magnetic Stand -96 (Ambion, Cat # AM10027) / or / similar for post PCR cleanup
- Heat block
The NEXTFLEX® Rapid Directional RNA Library Prep Kit contains enough material to prepare 8 or 48 RNA samples for Illumina® sequencing. The shelf life of all reagents is 12 months when stored properly. All components can safely be stored at -20°C.