Products

Buccal Swabs gDNA Extraction Kit(SpinColumn)

Packing Specification：

Product Introduction：
This kit adopts special imported DNA adsorption columns and a unique buffer system, specially designed for the isolation and purification of genomic DNA from buccal/pharyngeal swabs. After lysis and digestion of samples, DNA is selectively adsorbed onto the silica membrane of the spin column under high chaotropic salt conditions (Acryl Carrier is specially provided to easily capture trace nucleic acids from the system). Through a series of rapid washing and centrifugation steps, impurities such as salts, cellular metabolites, and proteins are removed. Finally, pure genomic DNA is eluted from the silica membrane using a low-salt Elution Buffer. The purified DNA is free of impurities and PCR inhibitors, and can be directly used for PCR analysis.

Product Features：
1.No need for toxic reagents such as phenol or ethanol precipitation steps.
2.Time-saving and simple; operation for a single sample can generally be completed within 20 minutes.
3.Equipped with Acryl Carrier for full collection of extremely trace DNA.
4.Multiple column washes ensure high purity. The extracted DNA has high purity and stable quality, suitable for various routine operations including PCR, enzyme digestion, sequencing, and Southern hybridization.

Application Scope：
Suitable for the isolation and purification of genomic DNA from buccal/pharyngeal swabs.

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!
Sample Collection:Take a sterile medical cotton swab (do not touch the absorbent cotton part with hands). Insert it into the mouth, press tightly against the inner cheek, and scrape back and forth 20 times (rotating the swab occasionally) to fully contact the oral mucosa.
Notes: Avoid touching the swab with hands. Rinse the mouth gently with water before collection. To prevent sample contamination by food or beverages, do not eat or drink within 30 minutes before sampling.

1.Use scissors to cut the cotton swab part from the stick and place it into a 2 mL centrifuge tube. Add 400 μL of Lysis Buffer ML.

2.Add 20 μL of Proteinase K (20 mg/mL) solution and immediately vortex to mix thoroughly.

3.Optional step (generally not required): Incubate at 56℃ for 1 hour, vortexing for 10 seconds every 10 minutes.
This step can be omitted under normal circumstances. Only attempt it if the extraction effect is poor.

4.Add 400 μL of Binding Buffer CB and immediately vortex to mix thoroughly. Incubate at 70℃ for 10 minutes.
If the number of cells on the swab is small, leading to low genomic DNA yield, add 4 μL of Acryl Carrier to 400 μL of Binding Buffer CB.

5.After cooling, add 200 μL of anhydrous ethanol and immediately vortex to mix thoroughly. Centrifuge briefly to remove droplets on the inner wall of the tube cap and collect all liquid at the bottom of the tube.
If the ambient temperature exceeds 25℃, pre-cool the anhydrous ethanol on ice before adding.

6.Load the mixture into a Spin AC column (placed in a collection tube). Centrifuge at 12,000 rpm for 30-60 seconds and discard the waste liquid in the collection tube.
The cotton of the swab may still absorb some liquid. To improve yield, squeeze out the liquid with clean tweezers before discarding the cotton to reduce liquid residue.

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

8.Add 500 μL of Wash Buffer WB (ensure anhydrous ethanol has been added!), centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

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

10.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).

11.Remove the Spin AC column and place it into a clean centrifuge tube. Add 20-50 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 1 minute, then centrifuge at 12,000 rpm for 1 minute. Transfer the eluted solution back into the spin column, incubate at room temperature for 1 minute, 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 20 μL; volumes smaller than this reduce elution efficiency and DNA yield).

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