KAPA HyperPrep Kit

Material Number: N/A

The streamlined, one-tube KAPA Hyper Prep protocol offers fast Library Prep turnaround time, with minimal hands-on time.

KAPA HyperPrep Kit

The KAPA HyperPrep Kit provides a versatile, streamlined protocol for the rapid construction of libraries for Illumina sequencing from fragmented, double-stranded DNA (dsDNA). The chemistry and streamlined, one-tube protocol improves the efficiency and consistency of library construction across a wide range of sample types and inputs.

The workflow combines enzymatic steps and employs minimal bead-based cleanups, thereby reducing sample handling and overall library preparation time to 2 – 3 hrs. The kit contains all of the enzymes and reaction buffers required for:

  1. End repair and A-tailing, which produces end-repaired, 5′-phosphorylated, 3′-dA-tailed dsDNA fragments;
  2. Adapter ligation, during which dsDNA adapters with 3′-dTMP overhangs are ligated to 3′-dA-tailed molecules;
  3. Library amplification (optional), which employs highf idelity, low-bias PCR to amplify library fragments carrying appropriate adapter sequences on both ends.

The kit provides a single, concentrated buffer and a single enzyme mixture for each of the two library construction steps. This offers the best combination of product stability, convenience and efficiency. Adapters and beads required for cleanups after adapter ligation and library amplification are not included. KAPA cleanup beads and KAPA Adapters are sold separately

The kit provides KAPA Pure Beads for reaction cleanups, along with all of the enzymes and buffers required for cDNA synthesis, library construction and amplification, but does not include RNA or adapters. KAPA Adapters are sold separately.

Reaction buffers are supplied in convenient formats comprising all of the required reaction components. This minimizes the risk of RNase contamination, ensures consistent and homogenous reaction composition, and improves uniformity among replicate samples. Similarly, a single enzyme mixture is provided for each step of the library construction process, reducing the number of pipetting steps.

In order to maximize sequence coverage uniformity and to maintain relative transcript abundance, it is critical that library amplification bias be kept to a minimum. KAPA HiFi DNA Polymerase has been designed for low-bias, high-fidelity PCR, and is the polymerase of choice for NGS library amplification[1,2,3,4]. The KAPA RNA HyperPrep Kit includes KAPA HiFi HotStart ReadyMix (2X) and Library Amplification Primer Mix (10X) for library amplification.

Product Applications

KAPA HyperPrep Kits are ideally suited for low- and high-throughput NGS library construction workflows that require end repair, A-tailing, adapter ligation and library amplification (optional). Kits are designed for library construction from a wide range of sample types and inputs (1 ng – 1 µg), and are compatible with complex genomic DNA, cell-free/circulating tumor DNA and lowquality DNA such as FFPE samples. For small genomes, cell-free/circulating tumor DNA and lower complexity samples such as ChIP DNA, amplicons or cDNA (for RNAseq), library construction from lower inputs (~100 pg or more) may be attempted. The protocol is automation-friendly and may be incorporated into workflows for a wide range of NGS applications, including:

  • whole-genome, shotgun sequencing
  • whole exome or targeted sequencing, using SeqCap EZ, Agilent SureSelect, Illumina TruSeq, or IDT xGen Lockdown Probes, or other hybridization capture systems
  • ChIP-seq
  • RNA-seq (starting with cDNA)
  • Methyl-seq (in combination with KAPA HiFi HotStart Uracil+ ReadyMix for library amplification).

 

1. Oyola, S.O., et al., BMC Genomics 13, 1 (2012).
2. Quail, M.A., et al., Nature Methods 9, 10 – 11 (2012).
3. Quail, M.A., et al., BMC Genomics 13, 341 (2012).
4. Ross, M.G., et al., Genome Biology 14, R51 (2013).

 

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The KAPA HyperPrep Presentations are:

KAPA HyperPrep Kit – 8 rxn | 7962312001 – KK8500
KAPA HyperPrep Kit – 24 rxn | 7962347001- KK8502
KAPA HyperPrep Kit – 96 rxm | 7962363001- KK8504

 

 

 

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8rxn, 24rxn, 96rxn