• DpnI

Product Application: Human iPS Cell Chemical Reprogramming Kit 2.0 (2nd Generation)

Primary Application:

Generation of Human Chemically Induced Pluripotent Stem Cells (hCiPSCs): This kit enables the efficient and stable reprogramming of human somatic cells, such as mesenchymal cells or fibroblasts derived from adipose tissue, skin, umbilical cord, and other tissues, into hCiPSCs.

Specific Uses:

  • Disease Modeling: hCiPSCs generated with this kit can be differentiated into various cell types to model human diseases, study disease mechanisms, and identify potential therapeutic targets.
  • Drug Discovery and Development: hCiPSC-derived cells provide a human-relevant platform for drug screening, toxicity testing, and evaluating the efficacy of novel therapeutic compounds.1
  • Regenerative Medicine Research: The hCiPSCs produced are a valuable source for generating replacement cells and tissues for potential therapeutic applications in regenerative medicine and cell therapy.2
  • Basic Research in Cell Biology and Stem Cell Science: Researchers can use this kit to investigate the fundamental mechanisms of cellular reprogramming, pluripotency, and epigenetic regulation in human cells.
  • Development of Personalized Medicine Approaches: The ability to efficiently reprogram somatic cells from individuals with diverse genetic backgrounds allows for the generation of patient-specific iPSCs for personalized disease modeling and therapeutic development.

Key Features and Benefits:

  • Fast and Efficient: The optimized system reduces the time required for induction to under 16 days, with high reprogramming efficiency.
  • Broad Applicability: Suitable for somatic cells from individuals of different genetic backgrounds and ages, ensuring robust and stable reprogramming.
  • Safe and Reliable: Chemical small molecules are used, eliminating the risks associated with genetic modification and potential immunogenicity.3
  • Simple Operation: The reprogramming process is conducted through straightforward medium exchange steps.
  • Standardized Production: The kit contains well-defined, serum- and feeder-free components, facilitating consistent and reproducible results.4
  • Non-Integrating: The chemical method avoids integration into the genome, enhancing the safety profile of the generated hCiPSCs.5

In summary, the BeiCell™ Human Chemical Reprogramming Kit (2nd Generation) provides a powerful and advanced tool for researchers and scientists seeking to generate high-quality human iPSCs for a wide array of applications in basic research, drug discovery, disease modeling, and regenerative medicine, with enhanced speed, efficiency, and safety.

Welcome to the documentation download page. Below you'll find important documents related to our products, including Certificates of Analysis (COA), Data Sheets, Material Safety Data Sheets (MSDS), and more. These documents provide essential information for product use, safety, and technical specifications.

Please note: The Certificate of Analysis (CoA) for this product is unavailable. You can download the Data Sheet and MSDS below. The User Manual is available upon request.

DpnI (Type IIM Restriction Enzyme)

REF: EG15585S

Recognition & Cleavage

Recognition site: 5'-GAm6TC-3'

Cleavage pattern:

5' ... G Am6T C ... 3'
3' ... C TAm6 G ... 5'
      

Type: IIM (recognizes & cleaves methylated DNA) · Common use: Removal of Dam-methylated parental template after PCR

Isoschizomer: MalI (methylation sensitivities may differ)

Recommended Reaction

  • Buffer: 1× CutOne® or 1× CutOne® Color Buffer
  • Incubation: 37 °C
  • Typical load: 1 µl enzyme per 20 µl reaction (≤1 µg plasmid DNA)
  • PCR template cleanup: Add 1 µl DpnI to 50 µl PCR, 37 °C 60 min; heat-inactivate 80 °C 20 min

CutOne® Color Buffer includes density reagent and tracking dyes for direct gel loading.

Quick Protocol (DNA Digestion)

  1. On ice, assemble 20 µl: ddH₂O up to 20 µl, 2 µl 10× CutOne® (or Color), ≤1 µg DNA, 1 µl DpnI (20 U/µl).
  2. Mix gently, quick-spin.
  3. Incubate at 37 °C for 15 min (plasmid/PCR); 30–60 min for genomic DNA.
  4. (Optional) Inactivate at 80 °C for 20 min or load gel directly if Color Buffer is used.

Notes

  • Keep total enzyme volume ≤10% of reaction.
  • For multiple digests, use 1 µl of each enzyme; if temperatures differ, start lower, then shift higher.
  • Increase incubation time if total volume >20 µl.

Quality Control

  • Functional test: 1 µg pUC19 + 1 µl DpnI → complete digest in 15 min at 37 °C.
  • Prolonged incubation/star activity: 3 h at 37 °C shows no nonspecific degradation (longer may result in star activity).
  • Non-specific endonuclease: <10% nicked/linearized after 4 h at 37 °C.

Methylation Effects

Methylase Effect
Dam No effect
Dcm No effect
CpG Impaired
EcoKI No effect
EcoBI Impaired

Recognition Sites in Common DNAs

DNA # Sites DNA # Sites
λDNA 116 ΦX174 0
pBR322 22 pUC57 15
pUC18/19 15 SV40 8
M13mp18/19 7 Adeno2 87

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DpnI

  • Brand: Best Enzymes
  • Product CAT#: EG15585S
  • Availability: In Stock

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Tags: Restriction Enzyme