Products

Processed Food gDNA Extraction Kit(SpinColumn)

Packing Specification：

Product Introduction:
The unique combination of Binding Buffer and Proteinase K rapidly lyses the material and inactivates intracellular nucleases. Genomic DNA is then selectively adsorbed onto the silica matrix membrane of the spin column under high chaotropic salt conditions. Through a series of rapid washing-centrifugation steps, Inhibitor Removal Buffer and Washing Buffer remove impurities such as cellular metabolites and proteins. Finally, pure genomic DNA is eluted from the silica matrix membrane using low-salt Elution Buffer.

Product Features:
1.The silica matrix membranes in the spin columns are all high-quality special adsorption membranes, with minimal variation in adsorption capacity between columns.
2.No toxic reagents such as phenol are required, nor are steps like ethanol precipitation.
3.Fast and simple: operation for a single sample can usually be completed within 30 minutes.
4.Multiple column washing steps ensure high purity. The typical OD260/OD280 ratio ranges from 1.7 to 1.9, and the DNA length can reach 30kb - 50kb, which can be directly used for PCR, Southern-blot, and various restriction enzyme digestion reactions.

Application Scope:
Suitable for rapid extraction of genomic DNA from various processed foods.

Operation Steps:
Tips:
Before first use, add the specified amount of absolute ethanol to Washing Buffer WB, mix thoroughly. After addition, mark the corresponding box to indicate ethanol has been added to avoid repeated additions!
Pretreat the adsorption column with Equilibration Buffer as a mandatory step; refer to "About the Use of Equilibration Buffer" above for specific methods.

1.Take 50 mg of sample (e.g., milk powder, biscuits, and other foods) and place it into a 1.5 mL centrifuge tube. Then add 200 μL of Buffer BB to resuspend.
If the subsequent extraction effect is unsatisfactory, the sample amount can be doubled to 100-200 mg, and the corresponding reagents should also be doubled proportionally.

2.Add 20 μL of Proteinase K (20mg/mL) solution, mix thoroughly, and incubate in a 65℃ water bath for 30 minutes.

3.Add 220 μL of Binding Buffer CB, mix thoroughly, and incubate in a 65℃ water bath for 10 minutes.

4.Centrifuge at 13,000 rpm for 3-5 minutes, aspirate the supernatant (approximately 400 μL) into a new centrifuge tube.

5.Add absolute ethanol equal to half the volume of the supernatant (about 200 μL), immediately vortex thoroughly to mix. Flocculent precipitation may occur at this step.

6.Transfer the mixture from the previous step (including any possible precipitation) to a Spin Column AC (placed in a collection tube), centrifuge at 13,000 rpm for 30-60 seconds, and discard the waste liquid in the collection tube.

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

8.Add 600 μL of Washing Buffer WB (check if absolute ethanol has been added!), centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

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

10.Place the Spin Column AC back into the empty collection tube, centrifuge at 13,000 rpm for 2 minutes to completely remove residual Washing Buffer (residual ethanol may inhibit downstream reactions).

11.Transfer the Spin Column AC to a clean centrifuge tube. Add 100 μL of Elution Buffer EB (pre-warming 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. If higher DNA concentration is required, the elution volume can be appropriately reduced, but the minimum volume should not be less than 50 μL (excessively small volumes reduce elution efficiency and DNA yield).

12.DNA can be stored at 2-8℃ for short-term use, or at -20℃ for long-term storage.