FAST & EASY DNA LIBRARY PREPARATION
NEXTFLEX® Rapid XP DNA-Seq Kit for Illumina® Platforms
- Reliable: Consistent yields and high-quality sequencing metrics
- Convenient: Kit includes fragmentation enzyme and cleanup/size selection beads
- Streamlined: Single step for fragmentation, end-repair, and adenylation with minimal hands-on time
- Flexible: Chemistry optimized with NEXTFLEX® color-balanced barcodes that allow a wide range of multiplexing (2 up to 384 samples in one run)
- Efficient: Automated on the PerkinElmer Sciclone® G3 and Zephyr® G3 NGS workstation
For research use only. Not for use in diagnostic procedures.
Streamlined Enzymatic Fragmentation and Library Prep Workflow for Illumina® Sequencing
The NEXTFLEX® Rapid XP DNA-Seq kit combines enzymatic fragmentation with end-repair and A-tailing in one reaction to create a highly efficient first step in library generation, which is followed by ligation and PCR (optional for PCR-free workflows). This effective one-tube workflow produces DNA-seq libraries with consistent library size, high yield, low GC-bias, and high coverage.
The kit includes all the required reagents for fragmentation, library prep, and magnetic bead-based cleanup, and it is compatible with your NEXTFLEX® barcodes of choice. This DNA-seq library prep kit is highly flexible in terms of sample requirements, accommodating a range of sample types and sample input amounts from 1 ng to 1 µg to quickly and efficiently generate high-quality libraries for Illumina® sequencing.
Figure 1: Reliable performance of NEXTFLEX® Rapid XP DNA-seq fragmentation enzyme. DNA is fragmented more evenly and into smaller sizes using the NEXTFLEX® Rapid XP DNA-seq kit than Competitor K’s kit. Each dot indicates each library peak size. (A) Libraries were prepared from 1 µg of DNA with 7 minutes of fragmentation with 2 PCR cycles: NEXTFLEX® Rapid XP DNA-seq kit (484 ± 52 bp, 11 %, n=17), Competitor K’s kit (1266 ± 451 bp, 36 %, n=17). (B) Libraries were prepared from 100 ng of DNA with 8 minutes of fragmentation with 5 PCR cycles: NEXTFLEX® Rapid XP DNA-seq kit (457 ± 12 bp, 3 %, n=18), Competitor K’s kit (730 ± 119 bp, 16 %, n=17). (C) Libraries were prepared from 1 ng of DNA with 15 minutes of fragmentation with 12 PCR cycles: NEXTFLEX® Rapid XP DNA-seq kit (448 ± 54 bp, 12 %, n=17), Competitor K’s kit (628 ± 160 bp, 25 %, n=17). All different input libraries were generated from 6 different commercially available DNA samples. (mean ± standard deviation, coefficient of variation)
Flexible Multiplexing Options
We offer a broad range of NEXTFLEX® adapters adapters that improve multiplexing capabilities and offer a solution to flexible experimental setups for both low-level and high-level multiplexing needs. The NEXTFLEX® Rapid XP DNA-seq kit is compatible with NEXTFLEX® DNA Barcodes, NEXTFLEX-96™ DNA Barcodes, NEXTFLEX® ChIP-Seq Barcodes, and NEXTFLEX-96™ ChIP-Seq Barcodes, and and NEXTFLEX-HT™ barcodes. For those high throughput users interested in dual index barcodes, the reagents are also compatible with the NEXTFLEX® Unique Dual Index Barcodes and the NEXTFLEX® Dual-Indexed barcodes.
For more information about barcode options compatible with the NEXTFLEX® Rapid XP DNA-seq kit, please contact NGS@perkinelmer.com.
The NEXTFLEX® Rapid XP DNA-seq kit is designed to function with multiple liquid handlers, including the PerkinElmer Sciclone® and the Zephyr® G3 NGS workstations. For more information, or to obtain a program, contact NGS@perkinelmer.com.
For availability of custom format and bulk packaging for larger volume requirements, please contact NGS@perkinelmer.com for further information.
Tech Tips – When to Size Select
Size selection is a critical issue in NGS library preparation. Our blog post, Tech Tips – When to Size Select, addresses a number of factors to consider before determining a size selection strategy.
- NEXTFLEX® Fragmentation Buffer
- NEXTFLEX® Fragmentation Enzyme Mix
- NEXTFLEX® Ligase Buffer Mix XP
- NEXTFLEX® Ligase Enzyme XP
- NEXTFLEX® PCR Master Mix XP
- NEXTFLEX® Primer Mix XP
- NEXTFLEX® Cleanup Beads XP
- Nuclease-free Water
- Resuspension Buffer
REQUIRED MATERIALS NOT PROVIDED
- 1 ng – 1 µg of DNA in up to 34 µL nuclease-free water.
- NEXTFLEX® barcodes
- Ethanol 80% (room temperature)
- 96 well PCR Plate Non-skirted (Phenix® Research, Cat # MPS-499) or similar
- 96 well Library Storage and Pooling Plate (Thermo Fisher® Scientific, Cat # AB-0765) or similar
- Adhesive PCR Plate Seal (Bio-Rad®, Cat # MSB1001)
- Magnetic Stand -96 (Thermo Fisher® Scientific, Cat # AM10027) or similar
- 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
- Nuclease-free barrier pipette tips
Figure 2: High yield obtained for libraries prepared using the NEXTFLEX® Rapid XP DNA-seq kit. Libraries were prepared using both NEXTFLEX® Rapid XP DNA-seq kit and Competitor K’s kit. (A) Libraries were prepared from 1 µg of DNA with 7 minutes of fragmentation with 2 PCR cycles. (B) Libraries were prepared from 100 ng of DNA with 8 minutes of fragmentation with 5 PCR cycles. (C) Libraries were prepared from 1 ng of DNA with 15 minutes of fragmentation with 12 PCR cycles.
Figure 3. Libraries prepared with the NEXTFLEX® Rapid XP DNA-seq kit show less or comparable GC-bias than libraries prepared with Competitor K’s kit. Picard GC bias analysis, which show read coverage over the indicated genome with respect to GC content, is shown as a fraction of normalized coverage (blue) calculated in 100 bp windows (red) and plotted against the left y-axis. Mean base quality at each window (green) is calculated and plotted against the right y-axis.
Figure 4. Libraries prepared with the NEXTFLEX® Rapid XP DNA-seq kit show higher or comparable genome coverage compared to libraries prepared using Competitor K’s kit. The percentage of genome bases covered at a minimum depth of 1X, 5X, and 10X of S. cerevisiae, E. coli, and B. pertussis.
Figure 5. Libraries prepared with the NEXTFLEX® Rapid XP DNA-seq kit resulted in better or comparable data with respect to mapping rate and duplication rate, compared to libraries prepared using Competitor K’s kit. (A) Mapping rates represented as the percentage of reads which aligned to the indicated genome. (B) Duplication rates represented as the percentage of reads predicted to be PCR duplicates by Picard MarkDuplicates.
The NEXTFLEX® Rapid DNA-Seq kit contains enough library prep reagents and cleanup beads to prepare 48 DNA samples for Illumina® sequencing. The shelf life of all reagents is at least 6 months when stored properly. The Nuclease-free Water and Resuspension Buffer can be stored at room temperature. The NEXTFLEX® Cleanup Beads XP should be stored at 4°C, and all other components should be stored at -20°C. The NEXTFLEX® Cleanup beads XP ships at room temperature, while other components ship on dry ice.