/ 1 / www.apexbt.com Protocol Cat. No. K1057 Tn5 DNA Library Prep Kit for Illumina (for 50 ng DNA) Introduction Tn5 DNA Library Prep Kit for Illumina (for 50 ng DNA) is an optimized genomic library construction kit for the Illumina high-throughput sequencing platform, which is suitable for the amount of 50 ng input DNA. Traditional library construction generally includes multi-step operations such as DNA fragmentation, end repair and linker ligation, while our kit adopts Tn5 transposase method which can be completed in one-step reaction. Our kits are easy to operate and greatly reduce library construction time, and significantly reducing the demanded amount of the initial DNA. The Transposase Mix and buffers provided by the kit are subject to rigorous quality control and functional verification, so as to achieve high fidelity, stability and repeatability. Components and Storage 24 rxns 96 rxns Components Transposase Mix 120 μl 480 μl 2X Tagmentation Buffer 240 μl 960 μl Amplify Enzyme 24 μl 96 μl 5X Amplify Buffer 240 μl 960 μl dNTP Mixture 96 μl 384 μl P5 60 μl 240 μl P7 60 μl 240 μl Control DNA 10 μl 10 μl Store Transposase Mix at 4°C and other components at –20C. *Note: Control DNA, Escherichia coli Genomic DNA, 50 ng/μl. Materials Not Supplied 1. Index Kit 1 for Illumina (APExBIO, Cat. No. K1058) or Index Kit 2 for Illumina (APExBIO, Cat. No. K1059) 2. Beads: Recommend DNA Clean and Selection Beads (APExBIO, Cat. No. K1060) or Agencourt AMPure XP, (BECKMAN, Cat. No. A63880-A63882)
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Protocol
Cat. No. K1057
Tn5 DNA Library Prep Kit for Illumina (for 50 ng DNA)
Introduction
Tn5 DNA Library Prep Kit for Illumina (for 50 ng DNA) is an optimized genomic library construction kit for the
Illumina high-throughput sequencing platform, which is suitable for the amount of 50 ng input DNA. Traditional
library construction generally includes multi-step operations such as DNA fragmentation, end repair and linker
ligation, while our kit adopts Tn5 transposase method which can be completed in one-step reaction. Our kits are
easy to operate and greatly reduce library construction time, and significantly reducing the demanded amount of
the initial DNA.
The Transposase Mix and buffers provided by the kit are subject to rigorous quality control and functional
verification, so as to achieve high fidelity, stability and repeatability.
Components and Storage
24 rxns 96 rxns Components
Transposase Mix 120 μl 480 μl
2X Tagmentation Buffer 240 μl 960 μl
Amplify Enzyme 24 μl 96 μl
5X Amplify Buffer 240 μl 960 μl
dNTP Mixture 96 μl 384 μl
P5 60 μl 240 μl
P7 60 μl 240 μl
Control DNA 10 μl 10 μl
Store Transposase Mix at 4°C and other components at –20C.
*Note: Control DNA, Escherichia coli Genomic DNA, 50 ng/μl.
Materials Not Supplied
1. Index Kit 1 for Illumina (APExBIO, Cat. No. K1058) or Index Kit 2 for Illumina (APExBIO, Cat. No. K1059)
2. Beads: Recommend DNA Clean and Selection Beads (APExBIO, Cat. No. K1060) or Agencourt AMPure XP,
(BECKMAN, Cat. No. A63880-A63882)
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3. Ethanol (100%)
4. Nuclease-free Water
Library Structure and Mechanism of Library Preparation
1. Mechanism of Library Preparation
Transposase Mix contains Tn5 transposase and two kinds of adapters (Adapter 1 and Adapter 2) with equal molar.
DNA fragmentation and end-linking were performed by incubating Transposase Mix with input DNA at 55C for 10
min. The tagged fragments can be further amplified with two pairs of primers, N5 (N5XX) and N7 (N7XX) and P5
and P7 (PCR Primer Mix), and the amplified product is subjected to size selection and purification to obtain the
DNA library for sequencing on the Illumina platform. The mechanism is shown in Figure 1: