LOW INPUT LIBRARY PREPARATION FOR NIMBLEGEN® SEQCAP® EZ LIBRARY CONSTRUCTION & ILLUMINA® SEQUENCING
NEXTFLEX® Pre-Capture Combo DNA Library Prep Kit for SeqCap® EZ Target Capture
- Complete kit – Contains library prep, barcodes and barcode blockers compatible with SeqCap® EZ target capture
- Low input – Only 10 ng of input DNA required
- Optimized – Offers larger number of on-target reads
- Multiplex up to 96 samples with single-index barcodes
- Reduces the cost of target capture
- Compatible with Illumina® sequencing platforms
For research use only. Not for use in diagnostic procedures.
24 BARCODES | 48 RXNS
48 BARCODES | 96 RXNS
96 BARCODES | 192 RXNS
Deep Multiplexing for Both Exome & DNA Target Capture
The NEXTFLEX® Rapid Pre-Capture Combo Kit is a complete low-input library prep solution designed for DNA library preparation, multiplexing, and barcode blocking, upstream of SeqCap EZ Library target enrichment. This kit facilitates deep multiplexing for both exome target capture and DNA target capture. The NEXTFLEX® Rapid PreCapture Combo Kit is designed to prepare single read or paired-end libraries for DNA and exome target capture and Illumina® sequencing. This kit features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads.
Automation for Library Prep for Target Capture
The NEXTFLEX® Rapid Pre-Capture Combo Kit is compatible with Beckman Biomek® FX and Biomek® FXP Laboratory Automation Workstations. A program is currently being validated for use with the Sciclone® NGS, Sciclone® NGSx and Sciclone® G3 workstations. For more information contact Bioo.NGS@perkinelmer.com.
Multiplex up to 96 Samples with SeqCap® Target Capture
The NEXTFLEX® DNA Barcodes and NEXTFLEX® DNA Barcode Blockers for SeqCap® are included in the NEXTFLEX® Rapid Pre-Capture Combo Kit and are also available separately. Our white paper written in conjunction with Roche®, An Integrated Solution to Simplify Library Preparation and Multiplexing for NimbleGen® Sequence Capture, demonstrates the efficacy of this kit when used with the NimbleGen® SeqCap® EZ Library system for library prep in preparation for Illumina® sequencing.
Achieving High Coverage & Yield from GC & AT Rich Genomes
The PCR amplification step in library preparation introduces biases including uneven coverage of regions with extreme base composition, increased numbers of duplicate fragments, decreased mapping quality and poor variant calling. In our white paper, Achieving High Coverage and Yield from GC and AT Rich Genomes, we show that the NEXTFLEX® PCR polymerase contains a highly optimized and robust enzyme that exhibits minimal GC bias and produces uniform coverage of difficult to sequence genomes.
- NEXTFLEX® End-Repair & Adenylation Buffer Mix
- NEXTFLEX® End-Repair & Adenylation Enzyme Mix
- NEXTFLEX® Ligase Enzyme Mix
- NEXTFLEX® DNA Barcode Adapter 1 – 24 (25 µM)
- NEXTFLEX® PCR Master Mix
- NEXTFLEX® Primer Mix (12.5 µM)
- NEXTFLEX® INV-HE Index 1 – 24 (250 µM)
- NEXTFLEX® HE Universal Oligo 1 (500 µM)
- NEXTFLEX® LM-PCR Oligo 1 (100 µM)
- NEXTFLEX® LM-PCR Oligo 2 (100 µM)
- NEXTFLEX® LM-PCR Master Mix
- Nuclease-free Water
- Resuspension Buffer
REQUIRED MATERIALS NOT PROVIDED
- 10 ng – 1 µg fragmented genomic DNA in up to 32 µL nuclease-free water.
- NimbleGen SeqCap EZ Library and related consumables
- NimbleGen SeqCap EZ Hybridization and Wash Kit (Roche, Cat. # 05634261001, 24 reactions) and related consumables
- NimbleGen SeqCap Pure Capture Bead Kit (Roche, Cat. # 06977952001, 24 reactions) and related consumables
- For Human Studies: COT Human DNA (Roche, Cat. # 11581074001)
- For Non-Human Studies: Nimblegen SeqCap EZ Developer Reagent (Roche, Cat. #06684335001)
- Ethanol 100% (room temperature)
- Ethanol 80% (room temperature)
- 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) / or / similar
- 96 well Library Storage and Pooling Plate (Fisher Scientific, Cat # AB-0765) / 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
- Heat block
- Thermocycler capable of holding 47ºC for 72 hours
- 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
- Nuclease-free barrier pipette tips
- 1.5 mL nuclease-free microcentrifuge tubes
Publications that Cite Using the NEXTFLEX® Rapid Pre-Capture Combo Kit:
- Suren, H., et al. (2016) Exome capture from the spruce and pine giga-genomes. Molecular Ecology Resources Mol Ecol Resour. doi:10.1111/1755-0998.12570.
The NEXTFLEX® Rapid Pre-Capture Combo Kit contains DNA library preparation reagents to prepare 48, 96 or 192 genomic DNA or exome libraries for SeqCap® EZ target capture and downstream Illumina® sequencing. The shelf life of all reagents is 12 months when stored properly at -20°C. This kit is shipped on dry ice.