Oral Swab Genomic DNA Extraction Kit

1. Components of the reagent kit

2. ストレージ

This reagent kit should be stored at room temperature (15-25℃) and in dry conditions, with a shelf life of 12 月. The DNA extraction purification columns can be stored for 1 year in a cool and dry environment. Proteinase K and RNase A contain preservatives, allowing transportation at room temperature, but for long-term storage, they should be kept at -20℃.

3. Instructions for Using the Reagent Kit

3.1 This reagent kit is intended for molecular biology research and should not be used for disease diagnosis or treatment.

3.2 Some components in the reagent kit contain irritants. Protective measures such as wearing protective clothing and goggles are recommended.

3.3 During the usage of this reagent kit, a high-speed centrifuge, 水浴 (metal bath), vortex mixer, anhydrous ethanol, sterile deionized water, and EP tubes need to be prepared by the user.

4. Introduction to the Reagent Kit

The Oral Swab DNA精製 Kit provides a rapid and efficient method for purifying DNA from oral swabs, widely used for the extraction of epithelial cells such as oral epithelium.

The Oral Swab DNA Rapid Purification Kit can extract total DNA from oral epithelial cells within 30 分. The entire purification process does not require toxic reagents such as phenol-chloroform. The extracted DNA can be directly used for PCR, Southern blotting, and other applications.

5. Experimental Principles and Procedures

6. Extraction Process

Before Starting the Experiment:

あ. Reagent Buffer B and C may precipitate under low-temperature conditions. We recommend heating at 65℃for 5 分. After the precipitate dissolves, it can be used normally.

B. ご使用の前に, add the specified amount of anhydrous ethanol to Wash Buffer 1as indicated on the bottle label. Mark a check on the label to indicate the addition of anhydrous ethanol.

C. Elution Buffer is a0.1x TE solutioncontaining minimal amounts of EDTA. If EDTA might affect subsequent experiments, it is advisable to substitute Elution Buffer with sterile deionized water.

1. Sample Handling:
2. Sampling: Take a sterilized cotton swab, insert it into the mouth, rub against the inner cheek back and forth for more than 20 回.
3. Use scissors to cut the head of the cotton swab, place it into a 2 ml centrifuge tube, and add 400μl of 試薬 Buffer B.
4. 追加10μl Proteinase K (10 mg/ml) and 10μl RNase A (10 mg/ml), invert thoroughly, incubate at 65°C for 20 分, vortex for 10 seconds during incubation.
5. 追加 400μl of 試薬 Buffer Cto the lysate, vortex thoroughly.
6. 追加 200μl of pre-chilled absolute ethanol, よく混ぜます; precipitation may occur, but it will not affect subsequent experiments.
7. Transfer the obtained liquid into a DNA extraction and purification column (キット) (approximately 650~700μl each time), で遠心分離する >8,000 の回転数 1 分, discard the collected waste liquid, and reinsert the collection tube into the DNA extraction and purification column (キット) for the next step.
8. Place the DNA extraction and purification column (キット) into a collection tube, 追加 300μl of Wash Buffer 1, で遠心分離する >8,000 の回転数 1 分, discard the waste liquid, and reinsert the DNA extraction and purification column (キット) into the tube for the next step.

(注記: Ensure that absolute ethanol has been added to Wash Buffer 1.)

7. 追加 500μl of Wash Buffer 1to the DNA extraction and purification column (キット), で遠心分離する 14,000 rpm (20,000×g) のために 2 分, extend the centrifugation time appropriately to dry the membrane further.
8. Place the DNA extraction and purification column (キット) into a new centrifuge tube, open the lid, incubate at 65°C for 2 分; this step can be extended appropriately to evaporate ethanol as much as possible, preventing residual ethanol from affecting downstream experiments.
9. Elute 100μl of Elution Bufferonto the membrane, で遠心分離する 12,000 の回転数 2 分.

(注記: 1. Eluting DNA with 50μl of Elution Buffer can increase DNA concentration but decrease total DNA yield; 2. Eluted DNA in the eluate can be reapplied to the DNA extraction and purification column, centrifuge again at 12,000 の回転数 2 minutes to increase DNA yield.)