Epigenetics

The Hieff NGS dsDNA Methyl Library Prep Kit - 12214ES
The Hieff NGS dsDNA Methyl Library Prep Kit for Illumina is a double-stranded DNA methylation library preparation kit developed for the Illumina high-throughput sequencing platform. This kit utilizes appropriate methylated adapters for ligation, followed by treatment with bisulfite or enzyme conversion, and then a PCR amplification step to construct methylated libraries for sequencing on the Illumina platform. All components of the kit undergo quality control and functional validation to ensure stable library output, supporting various types of DNA samples including gDNA, cfDNA, and FFPE. The recommended input DNA amount for this kit is 10 ng to 1000 ng, and it is compatible with commonly used conversion protocols.

Packaging size: 8 T / 24 T / 96 T

Hieff Superfast DNA Methylation Bisulfite Kit - 12225ES
The Hieff Superfast DNA Methylation Bisulfite Kit (Column-based) rapidly converts unmethylated cytosines in DNA samples to uracil, while leaving methylated cytosines unchanged. During high-temperature bisulfite treatment, double-stranded DNA denatures into single strands. In the presence of HSO3-, cytosine residues undergo deamination and are converted into uracil, with methylated cytosines remaining unaltered. In subsequent PCR amplification, uracil is replaced by thymine (T). The conversion process takes only 5 minutes, accommodating DNA input ranging from 100 pg to 2 μg, and achieves a conversion efficiency of ≥99% for unmethylated cytosines. The converted DNA is suitable for downstream applications such as PCR amplification and NGS sequencing.

Packaging size: 10 T / 50 T

To ensure the success of downstream experiments, accurately quantify the total amount of input DNA during the conversion step. It is recommended to use Qubit 3.0/4.0 for DNA quantification, with an A260/A280 ratio between 1.7 and 1.9. The input DNA range should be between 100 pg and 2 μg, with an optimal range of 100 ng to 1 μg. Insufficient DNA input can hinder downstream detection, while excessive input may reduce recovery and conversion efficiency.

Features:

Low Input: Suitable for converting samples ranging from 100 pg to 2 μg
Short conversion time: approximately 5 minutes
Minimal Sample Damage: Maintaining good sample integrity post-conversion
High Conversion Efficiency: Conversion rate ≥99%,  high conversion rates in high-GC regions with a low false positive rate
Suitable for rare samples, such as single-cell DNA methylation conversion
Capable of methylation conversion of RNA samples

Hieff NGS DNA Spike in Mix for CUT&Tag - 12596ES
Hieff NGS DNA Spike-in Mix for CUT&Tag (5 ng/μL) is derived from three sequences of Lambda DNA from E. coli, with lengths of 230 bp, 250 bp, and 300 bp, and a molar concentration ratio of approximately 1:3:10. This standard is primarily used for normalization and quantitative analysis of CUT&Tag sequencing data under different treatment conditions or varying cell states. Add 1 μL per 100,000 cells, with adjustments possible based on the binding capacity and abundance of the target protein.

Packaging size: 48 T

Notes:

The DNA Spike-in Mix provided by this product contains three sequences derived from E. coli λDNA, with a concentration of 5 ng/μL and lengths of 230 bp, 250 bp, and 300 bp, in a molar concentration ratio of approximately 1:3:10. For experiments, it is recommended to dilute the DNA Spike-in Mix with TE buffer to a concentration of 5 pg/μL.
This product is primarily used for normalization and quantitative analysis of sequencing data under different treatment conditions or cell states and is an optional component.

Nanobody-Tn5 - 12595ES
Nanobody-Tn5 is a recombinant Tn5 transposase fused with nanobody which can specific bind with primary antibodies from two different species (mouse and rabbit).

Packaging size: 10 μL/ 80 μL/ 100 μL

IFU - using high-throughput sequencing library preparation as an example:

Adapter preparation - Reference oligo name and sequence of Illumina platform
Dissolve the Oligo A, Oligo B, and Oligo C to 200 μM
Prepare the reaction I mix by adding Oligo  B and Oligo C  in 10uL each
Prepare the reaction II mix by adding Oligo A-1 or A-2 or A-3（200 μM) and ligo C in 10uL each
Mix reaction 1 and 2 together, and briefly centrifuge to bring the solution back to the bottom of the tube. Place in the PCR instrument, set up the following program and run it: hot lid at 105°C,and then heat at 94℃ for 2 min. After the time was up, the reaction procedure was stopped without opening the hot lid.The PCR tube was naturally cooled in the PCR instrument for 2 h, and then placed at 4℃ for 5 min.
After the reaction, mix the equal volumes of Reaction 1 and Reaction 2. Named ad Adapter Mix and stored from – 25 to – 15℃.
Prepare transposon generation mix by adding Anti-mouse Tn5 or Anti-rabbit Tn5 (10uL), Adapter mix (50 μM) (0.5uL) and Assemble Buffer (0.5uL)
Reaction conditions: use the pipette to gently blow and mix well. Perform the reaction at 25℃ for 1 h (hot lid off). The products of reaction can be directly applied to the library preparation or stored from -25 to -15℃.
Note: Transposase is temperature-sensitive, so it is recommended to complete the adapter embedding as soon as possible. The generated transposon can be stored at-25~ -15℃, with a validity period of 1 year.

Concanavalin A-Coated Magnetic Beads - 19810ES
Concanavalin A (ConA) beads are superparamagnetic polymer microspheres that have been covalently coupled with Concanavalin A (a plant mannose/glucose-binding lectin isolated from the seeds of cereal plants)on their surface. These beads posessess several notable characteristics, including monodispersity and strong magnetic reactivity. When Ca2+ and Mg2+ ions are present, ConA magnetic beads can efficiently and rapidly separate and purify various biomolecules such as polysaccharides, glycoproteins, and glycolipids by utilizing the affinity between ConA globulin A and the terminal α-D-mannose and α-D-glucose groups.
ConA magnetic beads offer a convenient method to separate or fix cells, ensuring minimal cell loss during subsequent washing steps. Additionally, they can be employed for the collection and fixation of nuclei. Moreover, these beads find utility in innovative techniques such as CUT & Run and CUT & Tag, which are revolutionary approaches used in ChIP-seq experiments.
In summary, ConA magnetic beads are powerful tools for the efficient purification of biomolecules, convenient cell separation and fixation, collection of nuclei, and application in cutting-edge experimental techniques.

Packaging size: 200 μL /1 mL/5 mL/20 mL