Manual

Product Number:PCK452

Shipping and Storage

Storage at -20°C and transport on dry ice.

Components

Description

This kit is specially developed for the Illumina high-throughput sequencing platform. It provides the enzyme premix system and reaction buffer required for genomic DNA library construction, including all components except PCR primers. Compared with traditional library construction kits, this kit uses a novel transposase method for library construction, which can complete DNA fragmentation, end repair and adapter ligation in one simple enzymatic reaction, significantly reducing the amount of template used. reducing the experimental operation steps and shortening the library construction time; using high-fidelity DNA polymerase for library enrichment, unbiased PCR amplification, expanding the coverage area of the sequence, and efficiently preparing DNA for the Illumina next-generation sequencing platform library. This kit is suitable for starting template DNA input of 50 ng. All reagents in the kit have undergone strict quality control and functional verification to ensure the stability and reproducibility of library construction to the greatest extent.

Reagents to Be Supplied by User

1. Magnetic stand.
2. DNA purification and recovery kit
3. Library PCR primer kit and PCR thermal cycler
4. Absolute ethanol, deionized water (pH between 7.0-8.0).
5. Reaction tubes: It is recommended to use low adsorption PCR tubes and 1.5 ml centrifuge tubes.
6. Pipette tips: It is recommended to use high-quality filter tips to prevent contamination of kits and library samples.

Important Points Before Starting

1. Avoid repeated freezing and thawing of reagents.
2. The PCR products are easily contaminated 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 special pipette to regularly clean each experimental area.
3. Magnetic bead purification: The magnetic beads should be equilibrated to room temperature before use. All operations on 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 frosted. , Insufficient drying of magnetic beads will cause ethanol residue to affect subsequent experiments, and excessive drying of magnetic beads will affect DNA recovery efficiency.
4. This kit is suitable for the construction of human genomic DNA library. If the DNA sample is a PCR product, its length should be more than 500 bp. Since the transposase cannot act on the DNA end, it is recommended to extend the PCR product at both ends when preparing the PCR product. 50-100 bp to avoid low end sequencing coverage.

Sample preparation

1. DNA purity requirements: A260/A280 = 1.8-2.0.
2. Sample DNA: Dissolve in ultrapure water.
3. DNA quantification: Too much or too little DNA input will affect the quality of the library. It is recommended to use Nano to detect the purity of genomic DNA and then to use Qubit to detect the concentration of the genome. Each sample is measured 3 times and the average value is obtained (do not use Template quantification based on any absorbance measurement-based assay).