KAPA Low Throughput Library Preparation Kit PCR-free


KAPA low throughput Library Preparation Kit, PCR-free (illumina).



KAPA low throughput Library Preparation Kit, PCR-free (illumina).

The KAPA LTP Library Preparation Kit is designed for the construction of libraries for Illumina sequencing, starting from fragmented, double-stranded DNA (dsDNA). The kit contains all of the enzymes and reaction buffers required for the following steps of library construction:

1. end repair, to produce blunt-ended, 5′-phosphorylated dsDNA fragments;

2. A-tailing, during which dAMP is added to the 3′-ends of blunt-ended dsDNA library fragments;

3. adapter ligation, during which dsDNA adapters with 3′-dTMP overhangs are ligated to 3′-A-tailed library fragments; and

4. library amplification (optional), which employs highfidelity, low-bias PCR to amplify library fragments carrying appropriate adapter sequences on both ends.

Efficient, cost-effective reaction cleanups and higher library yields are achieved through implementation of the “with-bead” strategy developed at The Broad Institute of MIT and Harvard and Foundation Medicine.1 The kit includes PEG/NaCl Solution for this purpose. Adapters and beads required for cleanups after end repair and library amplification are not included. KAPA Pure Beads (KK8000, KK8001, KK8002) and KAPA Adapters are sold separately.

In order to maximize sequence coverage uniformity, it is critical to minimize library amplification bias. KAPA HiFi DNA Polymerase is designed for low-bias, highfidelity PCR, and is the reagent of choice for NGS library amplification.2,3,4,5 KAPA LTP Library Preparation Kits include KAPA HiFi HotStart ReadyMix (2X), a ready-touse PCR mix comprising all the components for library amplification—except primers and template. Kits also include Library Amplification Primer Mix (10X), designed for the high-efficiency amplification of Illumina libraries flanked by adapters containing the P5 and P7 flow cell sequences. Kits without the amplification module (KK8231 and KK8233) are available for PCR-free workflows. They may also be combined with KAPA HiFi Real-time Library Amplification Kits (KK2701 and KK2702), or with KAPA HiFi HotStart Uracil+ ReadyMix Kits (KK2801 and KK2802) for the amplification of bisulfite-converted libraries.

1. Fisher, S., et al., Genome Biology 12, R1 (2011). 2. Oyola, S.O., et al., BMC Genomics 13, 1 (2012). 3. Quail, M.A., et al., Nature Methods 9, 10 (2012). 4. Quail, M.A., et al., BMC Genomics 13, 341 (2012). 5. Ross, M.G., et al., Genome Biology 14, R51 (2013).