Manual

Product Number: PCK246

Shipping and Storage

Store at -20℃ and transport on dry ice.

Components

Component	PCK246 24rxns	PCK246 96rxns
TPS V5	168μl	672μl
5×FA Reaction Buffer	96μl	384μl
TS Buffer	72μl	288ml
2×PCR Mix	600μl	2×1.2μl

Note: This kit is suitable for constructing human genomic DNA libraries, with a starting template DNA input of 5ng. Our company also has NGS TPH DNA Library Preps Kit for lllumina (1ng) for 1 ng (PCK247). To obtain a high-quality library, it is recommended to use different reagent kits for different DNA starting quantities.

Description

This kit is a specialized kit developed specifically for the Illumina high-throughput sequencing platform, providing the enzyme premix system and reaction buffer required for constructing genomic DNA libraries, including all components except PCR primers. Compared with traditional library construction kits, this kit adopts a novel transposase method for library construction, which can complete DNA fragmentation, end repair, and connector connection reactions through a simple enzymatic reaction, significantly reducing the use of templates, reducing experimental steps, and shortening library construction time; The use of high fidelity DNA polymerase for library enrichment and preference free PCR amplification has expanded the coverage area of the sequence, enabling efficient preparation of DNA libraries for the Illumina second-generation sequencing platform. This reagent kit is suitable for a starting template DNA input of 5ng. All reagents in the reagent kit have undergone strict quality control and functional verification, ensuring the stability and repeatability of library construction to the greatest extent possible.

Features

1. DNA fragmentation and splicing are completed in one step.
2. Ultra fidelity amplification minimizes amplification preference to the greatest extent possible.

Self provided instruments, reagents, and consumables

1. Magnetic frame
2. DNA purification and recovery kit: It is recommended to use the Magnetic Bear DNA Purification Kit (for NGS Size Selection) (DNK2508).
3. Library PCR Primer Kit
4. Anhydrous ethanol, deionized water (pH between 7.0-8.0).
5. Reaction tube: It is recommended to use a low adsorption PCR tube and a 1.5 ml centrifuge tube.
6. Gun tip: It is recommended to use high-quality filtering gun tips to prevent contamination of reagent kits and library samples.

Preparation and important precautions before the experiment

1. Avoid repeated freezing and thawing of reagents.
2. PCR products are prone to contamination due to improper operation, resulting in inaccurate experimental results. It is recommended to isolate the PCR reaction system preparation area from the PCR product purification area, and use a dedicated pipette to regularly clean each experimental area.
3. Magnetic bead purification: Before use, the magnetic beads should be balanced to room temperature. All operations of the magnetic beads should be carried out at room temperature. 80% ethanol should be prepared and used immediately. After rinsing, the magnetic beads should be dried until the surface has no liquid reflection and is in a frosted state. Insufficient drying of the magnetic beads will have residual ethanol, which will affect subsequent experiments. Excessive drying of the magnetic beads will affect the efficiency of DNA recovery.
4. This kit is suitable for constructing human genomic DNA libraries. If the DNA sample is a PCR product, its length should be ensured to be greater than 500bp. As transposase cannot act on the end of DNA, it is recommended to extend the two ends of the PCR product by 50-100bp each when preparing the PCR product to avoid low end sequencing coverage.

Sample preparation

1. DNA purity requirement: A260/A280=1.8-2.0.
2. Sample DNA: Dissolved in ultrapure water.
3. DNA quantification: Excessive or insufficient DNA input can have an impact on the quality of the library. It is recommended to first use Nano to test the purity of genomic DNA, and then use Qubit to perform concentration testing on the genome (do not use any absorbance based measurement methods for template quantification).

Schematic diagram of DNA library construction process