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FFPE Tissue Kit

VAMNE MagUltra

 DNA FFPE Tissue Kit for DNA Extraction DM601

 

size

50 rxns

Cat. No.

DM601-01




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This kit employs a safe, non-toxic, and environmentally friendly deparaffinization solution along with an efficient lysis/de-crosslinking reagent, which can lyse and release trace DNA in paraffin embedded sections and tissues. The high-affinity magnetic beads used in the kit can adsorb nucleic acids in a high-salt buffer and release nucleic acids in a low-salt elution buffer, thereby achieving rapid separation and purification of nucleic acids. The obtained products has good integrity and amplifiability, suitable for downstream applications such as PCR, next-generation sequencing, hybridization capture, etc. This kit can be used with automated nucleic acid extraction instrument for high-throughput extraction.

  • Safe and non-toxic: Use eco-friendly and non-toxic dewaxing agents instead of traditional toxic xylene for dewaxing
  • High yield and quality: The extracted product has a high yield and purity, with no inhibition for downstream applications, 

and the extraction effect is stable and reliable.

Storage  :

BOX 1: Store at 2 ~ 8°C and ship on ice pack.

BOX 2: Store at 15 ~ 25°C and ship at room temperature.

Mechanism and workflow :

                                                                                FFPE Tissue Kit workflow

FFPE DNA Extraction Protocol – Magnetic Bead Method

Optimized DNA Extraction from Paraffin-Embedded and Formalin-Fixed Tissues

This optimized FFPE DNA extraction protocol is designed for efficient purification of high-quality genomic DNA from:

  • Paraffin sections (FFPE samples)
  • Paraffin-embedded tissue blocks
  • Formalin-fixed tissues

The method combines deparaffinization, proteinase K digestion, de-crosslinking, and magnetic bead-based nucleic acid purification, ensuring high yield and purity for downstream applications such as PCR, qPCR, NGS, and sequencing analysis.

Step 1 – Sample Pretreatment

A. DNA Extraction from Paraffin Sections

Use 0.5–5 paraffin sections (10 µm thickness, approx. 30 mm² tissue surface).

  1. Scrape tissue into a 1.5 ml microcentrifuge tube.
  2. Add 800 µl Deparaffinization Solution.
  3. Vortex 5 seconds vigorously.
  4. Incubate at 56°C for 5 minutes.
  5. Vortex 15 seconds.
  6. Proceed to lysis and nucleic acid binding.

This step ensures effective paraffin removal for optimal DNA recovery from FFPE tissue.

B. DNA Extraction from Paraffin-Embedded Tissue Blocks

  1. Scrape approximately 10 mg tissue into a 1.5 ml tube.
  2. Add 800 µl Deparaffinization Solution.
  3. Vortex 5 seconds.
  4. Incubate at 56°C for 5 minutes.
  5. Vortex 15 seconds.

Continue with the DNA lysis step.

C. DNA Isolation from Formalin-Fixed Tissue

  1. Cut ~10 mg tissue into small pieces.
  2. Add 500 µl PBS.
  3. Centrifuge at 12,000 rpm (13,500 × g) for 1 minute.
  4. Discard supernatant.
  5. Repeat washing step three times.

Proceed to lysis and DNA purification.

This process improves DNA extraction efficiency from formalin-fixed samples by removing residual fixatives.

Step 2 – Lysis, De-Crosslinking and Magnetic Bead DNA Binding

  1. Add 40 µl Proteinase K.
  2. Add 200 µl L/D Buffer.
  3. Incubate at 56°C for 60–180 minutes (complete digestion).
  4. Incubate at 90°C for 60 minutes (reverse formalin cross-linking).

Transfer approximately 220 µl digestion solution into a new tube.

⚠ Important: Avoid transferring the upper deparaffinization layer.

Optional RNA removal:

Add 2 µl RNase A (100 mg/ml), incubate 3–5 minutes at room temperature.

Magnetic Bead DNA Purification

  1. Add 400 µl FB Buffer.
  2. Add 300 µl magnetic beads.
  3. Incubate 3 minutes at room temperature.
  4. Place on magnetic rack for 2 minutes.
  5. Discard supernatant.

This ensures strong DNA binding to magnetic beads for high-purity extraction.

Step 3 – Washing and DNA Elution

Washing Steps

  • Add 500 µl WA Buffer → magnetic separation → discard supernatant.
  • Repeat once.
  • Add 500 µl 80% ethanol → magnetic separation → discard.
  • Repeat once.

Air dry 5–10 minutes.

DNA Elution

  1. Add 50–100 µl Elution Buffer.
  2. Incubate at 55°C for 5 minutes.
  3. Place on magnetic rack.
  4. Transfer purified DNA to new tube.
  5. Store at –20°C.

Applications

Purified DNA is suitable for:

  • PCR
  • qPCR
  • NGS library preparation
  • Mutation analysis
  • Forensic DNA testing