Oral Swab Genomic DNA Extraction Kit

1. 試薬キットのコンポーネント

仕様	50T	100T
猫. いいえ.	SN0233	SN0234
DNA抽出カラム (セット)	50 (セット)	100 (セット)
Reagent Buffer B	20 ミリリットル	2×20 ミリリットル
Reagent Buffer C	30 ミリリットル	2×30 ミリリットル
洗浄バッファー 1	15 ミリリットル	2 × 15 ミリリットル
溶出バッファー	20 ミリリットル	20 ミリリットル
プロテイナーゼK	1ミリリットル	1ミリリットル
RNase A	1ミリリットル	1ミリリットル
取扱説明書	1	1

 

2. ストレージ

この試薬キットは、室温で保管する必要があります (15-25℃) and in dry conditions, 貯蔵寿命があります 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, しかし、長期保管の場合, they should be kept at -20℃.

3. 試薬キットを使用するための指示

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 (メタルバス), 渦ミキサー, 無水エタノール, 滅菌脱イオン水, and EP tubes need to be prepared by the user.

4. 試薬キットの紹介

 

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. 精製プロセス全体では、フェノールクロロホルムなどの有毒試薬は必要ありません. The extracted DNA can be directly used for PCR, サザンブロット, その他のアプリケーション.

5. 実験原則と手順

6. 抽出プロセス

Before Starting the Experiment:

あ. Reagent Buffer B and C 低温条件下で沈殿する可能性があります. We recommend heating at 65℃for 5 分. After the precipitate dissolves, it can be used normally.

B. ご使用の前に, 指定された量の無水エタノールを追加します 洗浄バッファー 1as indicated on the bottle label. Mark a check on the label to indicate the addition of anhydrous ethanol.

C. 溶出バッファーはaです0.1X TEソリューションcontaining minimal amounts of EDTA. If EDTA might affect subsequent experiments, it is advisable to substitute Elution Buffer with sterile deionized water.

1. サンプル処理:
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遠心チューブ, and add 400μlの 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の 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, しかし、それは後続の実験に影響しません.
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 300洗浄バッファーμl 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) 次のステップのためにチューブに.

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

7. 追加 500洗浄バッファーμl 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) 新しい遠心チューブに, 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の溶出緩衝液膜に, 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, 再び遠心分離機 12,000 の回転数 2 minutes to increase DNA yield.)