Product Number: PCK48

Shipping and Storage

Box 1 -20℃ storage, dry ice transportation.

Box 2 2-8℃, transported in ice bags

Components

• Box 1

• Box 2

Description

The Universal DNA Library Kit (Illumina&MGI) is a second-generation sequencing enzyme digestion library kit developed for Illumina and MGI sequencing platforms. It includes three modules: DNA fragmentation/end repair/addition of A, adapter connection, and pre mixed enzymes required for library enrichment in library construction.By controlling the reaction time, sample DNA fragments from different sources such as 0.1ng-1μg genomic DNA, PCR amplification products, and FFPE can be reduced to small fragments, avoiding tedious ultrasound processes and instrument dependence.This kit simplifies the purification steps and shortens the library construction time. In the amplification module, high fidelity DNA polymerase is used for library enrichment, and PCR amplification is performed without preference, ensuring the accuracy of sequencing results.

Features

1. Accurate enzyme digestion, fragment end repair and A tube completion, without the need for purification and direct connection to the connector.
2. After connecting the joint, use matching magnetic beads for direct sorting (optional).
3. High fidelity enzymes for PCR enrichment and amplification minimize amplification preference.
4. Suitable for building libraries for different species such as humans, animals, plants, microorganisms, etc. The resulting library is applicable to various platforms such as Illumina and MGI.

Schematic diagram of DNA database construction process

DNA library construction process

Please read this operating instruction carefully before the experiment and choose the operating plan based on the type of sequencing platform used. Before the experiment starts, clarify the nucleic acid concentration, and the input amount of this reagent kit is 0.1ng-1μg. The recommended sample size is 1ng-500ng DNA. DNA solution does not contain chelating agents, if DNA dissolves in 1×TE or EDTA containing solutions, it is recommended to use magnetic beads for purification or add Neutralization Reagent.

Product Number: FER46

Shipping and Storage

Store at -20℃ and transport on dry ice.

Components

Description

Onestep DNA Frag And ER Reagent is a new generation of enzyme digestion repair reagents developed for Illumina and MGI high-throughput sequencing platforms. It can complete one-step fragmentation/final repair/addition of A. This product includes FER Enzyme Mix and FER Buffer, which can achieve fragmentation and end repair in one tube, avoiding cumbersome ultrasound processes and instrument dependence, and is easy to operate. It can achieve efficient and fast fragmentation, end repair, and dA tail addition for different DNA samples, and is applied to downstream library construction. It has the advantages of low preference, stable enzyme digestion repair effect, and higher library transformation efficiency. It can be applied to fragment, end repair, and A addition reactions of samples from different sources such as 0.1 ng-1ug genomic DNA, PCR amplification products, FFPE, etc.

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).

Product Number: PCK84

Shipping and Storage

-20 ℃, 12 months. If frequently used, it can be stored at 2-8 ℃ to avoid repeated freeze-thaw as much as possible.

Components

Description

This product uses the dye method (SYBR Green I) to perform real-time fluorescence quantitative PCR (qPCR) on the products after NGS library construction. The reagent kit provides the reaction mixture, DNA primer mixture, standards, and complete reagent system required for the qPCR process, making the operation simple and convenient.This kit uses a chemically modified new highly efficient hot start polymerase, and the activation of the enzyme requires incubation at 95℃ for 10 minutes. This product has strong specificity and high amplification efficiency, and can quickly and accurately quantify the concentration of the constructed library. Suitable for fluorescence quantitative PCR instruments that do not require ROX as a calibration dye, such as Roche LightCyler 480, Roche LightCyler 96, Bio radioCyleriQ, iQ5, CFX96.

ROX dye is used to correct the fluorescence signal error generated between wells in quantitative PCR instruments, and is generally used in Real Time PCR amplification instruments from companies such as ABI and Stratagene. The excitation optical systems of different instruments vary, so the concentration of ROX dye must be matched with the corresponding fluorescence quantitative PCR instrument.

Instruments that do not require ROX calibration:Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96 etc.

Instruments that require Low ROX calibration:ABI Prism7500/7500 Fast，QuantStudio® 3 System，QuantStudio®5 System，QuantStudio®6 Flex System，QuantStudio®7 Flex System，ViiA 7 system，Stratagene Mx3000/Mx3005P，Corbett Rotor Gene 3000 etc.

Instruments that require High ROX calibration:ABI Prism 7000/7300/7700/7900，Eppendorf，ABI StepOne/Step One Plus etc.

Note:The preparation methods for High Rox and Low Rox are described in protocol Step 2.

Application

This product is designed for the Illumina platform second-generation sequencing library concentration absolute quantification. The end of the library contains Illumina P5 and P7 chip binding sequences, with a length of no more than 1 kb and a concentration of no less than 0.002 pM. This product can be used for quantitative experiments. The qPCR Primer Mix provided by the kit contains the following two primer sequences:

1. Primer 1: 5′-AAT GAT ACG GCG ACC ACC GA-3′
2. Primer 2: 5′-CAA GCA GAA GAC GGC ATA CGA-3′

It is possible to confirm in advance whether the library can be amplified by the primer pair through the primer sequence.

Storage

Store at -20 ℃ and transport on dry ice.

Components

Description

This kit provides adapters and primers used in second-generation sequencing library construction, specifically designed for Illumina platform library construction. The index primer design can meet the needs of preparing multiple library samples for multiple high-throughput sequencing. The prepared library can be used for sequencing platforms such as Illumina GAIIx, HiSacnSQ, HiSeq 2500/2000/1000, and MiSeq sequencing.

Product Number: PCK85M

Storage

Store at -20 ℃ and transport on dry ice.

Components

Description

This kit is suitable for both Illumina and MGI sequencing platforms. It provides the premixed enzyme modules required for DNA end repair,5′-end phosphorylation modification,3′-end A addition and Adaptor ligation in DNA library preparation. It can be used with different sequencing platform adapters. Primer kits can prepare DNA into specific DNA libraries for Illumina or MGI sequencing platforms. Using high-fidelity DNA polymerase for library enrichment and unbiased PCR amplification,the coverage area of the sequence is expanded,and high-quality DNA libraries can be prepared. All reagents provided in the kit have undergone strict quality control and functional verification to ensure the stability of library preparation to the greatest extent.

Features

1. End repair and phosphorylation and adding A to complete in one step;
2. After the end repair without purification,directly add the Adaptor;
3. Ultra fidelity amplification,the maximum extent to reduce the amplification bias;
4. The library is suitable for multiple sequencing platforms:MGI sequencing platform:MGISEQ-2000, MGISEQ-200, BGISEQ-500 and other MGI sequencing platform lllumina GAllx, HiSacnSQ, HiSeq 250/2000/1000, MiSeqsequencing and other lllumina platform sequencers;
5. It is suitable for preparing gDNA and cDNA libraries after physical interruption.

Schematic diagram of DNA library construction process

Product Number: PCK85

Storage

Store at -20 ℃ and transport on dry ice.

Components

Description

This kit provides the enzyme premix system and reaction buffer required for DNA library construction, including all components except connectors and PCR primers, for the construction of Illumina second-generation sequencing platform DNA library. Compared with general library building methods, this kit is simple and convenient to operate, greatly shortening the library construction time. In addition, the kit uses high-fidelity DNA polymerase for library enrichment, with no preference for PCR amplification, expanding the coverage area of the sequence, and can efficiently prepare a DNA library for the Illumina second-generation sequencing platform. All reagents provided in the kit have undergone strict quality control and functional verification, ensuring the stability of library construction to the greatest extent possible.

Features

1. End leveling, phosphorylation, and addition of A are completed in one step.
2. No need for purification, add connectors directly.
3. Ultra fidelity amplification minimizes amplification preference.
4. Supports multiple samples, and the resulting library can be used for sequencing platforms such as Illumina GAIIx, HiSacnSQ, HiSeq 2500/2000/1000, and MiSeq sequencing.

Schematic diagram of DNA database construction process