Products

FFPE Tissue gDNA Extraction Kit(SpinColumn)

Packing Specification：

Product Introduction：
Formalin-fixed or paraffin-embedded tissues are rapidly lysed to release genomic DNA through the combined action of unique lysis buffer heat treatment and Proteinase K. Genomic DNA is then selectively adsorbed onto the silica membrane of the spin column under high chaotropic salt conditions. Through a series of rapid washing and centrifugation steps, Inhibitor Removal Buffer and Wash Buffer remove impurities such as cellular metabolites and proteins. Finally, pure genomic DNA is eluted from the silica membrane using a low-salt Elution Buffer.

Product Features：
1.The silica membrane in the spin column is a special adsorption membrane imported from a world-renowned company, ensuring minimal variation in adsorption capacity between columns and good reproducibility. It overcomes the drawback of unstable membrane quality in domestic kits.
2.No need for toxic reagents such as phenol or ethanol precipitation steps.
3.Fast and simple; operation for a single sample can generally be completed within 30 minutes.
4.Multiple column washes ensure high purity. The typical OD260/OD280 ratio is 1.7-1.9, and the DNA length can reach 30 kb - 50 kb, which can be directly used for PCR, Southern blot, and various restriction enzyme digestion reactions.

Application Scope：
Suitable for the rapid extraction of DNA from various formalin-fixed and paraffin-embedded tissues.

Operation Steps：
Tip: Before first use, add the specified amount of anhydrous ethanol to Wash Buffer WB, mix thoroughly, and mark the box to indicate ethanol has been added to avoid repeated addition!

1.Place tissue sections into a centrifuge tube and soak in xylene for dewaxing for approximately 30 minutes (adjust the time according to the section thickness).

2.Rehydrate the sections by sequentially soaking them in 100% ethanol/80% ethanol/60% ethanol/40% ethanol/deionized water for 10 seconds each. 
The sections should turn white when first immersed in 100% ethanol.

3.Under microscopic observation, use a blade to cut the target tissue for DNA extraction and place it into a pre-weighed 1.5 mL centrifuge tube. Weigh again to calculate the weight of the sectioned tissue.
4.Add 200 μL of Lysis Buffer FTL and 20 μL of Proteinase K solution (20 mg/mL) to 25-50 mg of tissue. Mix immediately and incubate in a 37℃ water bath overnight.

5.Add another 10 μL of Proteinase K solution (20 mg/mL), mix, and incubate in a 55℃ water bath for 1-2 hours.
No large tissue particles should be visible after this step.

6.Add 200 μL of Binding Buffer CB, immediately vortex for 20 seconds to mix thoroughly, and incubate in a 70℃ water bath for 10 minutes.

7.After cooling, add 100 μL of isopropanol, immediately vortex for 30 seconds to mix thoroughly (flocculent precipitation may occur).

8.Use a 1 mL pipette tip to aspirate the mixture and load it into a Spin AC column (placed in a collection tube). Centrifuge at 13,000 rpm for 60 seconds and discard the waste liquid in the collection tube.
If insoluble tissue debris clogs the pipette tip, gently rub the tip on absorbent paper to remove the debris. If only a small amount of mixture is aspirated, discard the tip and debris together to avoid clogging the spin column.

9.Add 500 μL of Inhibitor Removal Buffer IR, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

10.Add 600 μL of Wash Buffer WB (ensure anhydrous ethanol has been added!), centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

11.Add 600 μL of Wash Buffer WB, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

12.Place the Spin AC column back into the empty collection tube and centrifuge at 13,000 rpm for 2 minutes to thoroughly remove residual Wash Buffer (residual ethanol may inhibit downstream reactions).

13.Remove the Spin AC column and place it into a clean centrifuge tube. Add 100 μL of Elution Buffer EB (preheating the elution buffer in a 65-70℃ water bath beforehand improves elution efficiency) to the center of the adsorption membrane. Incubate at room temperature for 3-5 minutes, then centrifuge at 12,000 rpm for 1 minute. Transfer the eluted solution back into the spin column, incubate at room temperature for 2 minutes, and centrifuge at 12,000 rpm for 1 minute.
Larger elution volumes result in higher elution efficiency. For higher DNA concentration, reduce the elution volume (minimum 50 μL; volumes smaller than this reduce elution efficiency and DNA yield).

14.Store DNA at 2-8℃ for short-term use or -20℃ for long-term storage.