Oral Swab Genomic DNA Extraction Kit

1. 試劑盒的組成

2. 貯存

該試劑盒應在室溫下保存 (15-25℃) 並且在乾燥條件下, 保存期限為 12 月. DNA 萃取純化柱可保存 1 在涼爽乾燥的環境中度過一年. 蛋白酶 K 和 核糖核酸酶A 含防腐劑, 允許在室溫下運輸, 但為了長期儲存, 應保存在-20℃.

3. 試劑盒使用說明

3.1 本試劑盒適用於分子生物學研究，不得用於疾病診斷或治療.

3.2 試劑盒中部分成分含有刺激物. 建議採取防護措施，例如穿著防護衣和護目鏡.

3.3 本試劑盒使用過程中, 高速離心機, 水浴 (金屬浴), 渦旋混合器, 無水乙醇, 無菌去離子水, 和EP管需用戶自備.

4. 試劑盒簡介

The Oral Swab DNA純化 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 分分鐘. 整個純化過程不需要苯酚氯仿等有毒試劑. 萃取的DNA可直接用於 聚合酶鍊式反應, 南方印跡, 和其他應用.

5. 實驗原理和程序

6. 提取過程

開始實驗前:

A. Reagent Buffer B and C 低溫條件下可能會沉澱. 我們建議在 65℃ 加熱 5 分分鐘. 沉澱溶解後, 可以正常使用.

乙. 使用前, 加入規定量的無水乙醇 洗滌緩衝液 1如瓶子標籤上所示. 在標籤上打勾，表示添加了無水乙醇.

C. 洗脫緩衝液是0.1x TE 解決方案含有極少量的 EDTA. EDTA是否可能影響後續實驗, 建議用無菌去離子水取代洗脫緩衝液.

1. 樣品處理:
2. Sampling: Take a sterilized cotton swab, insert it into the mouth, rub against the inner cheek back and forth for more than 20 次.
3. Use scissors to cut the head of the cotton swab, place it into a 2 毫升離心管, and add 400微升的 試劑 Buffer B.
4. 添加10μl Proteinase K (10 毫克/毫升) and 10μl RNase A (10 毫克/毫升), invert thoroughly, incubate at 65°C for 20 分分鐘, vortex for 10 seconds during incubation.
5. 添加 400微升的 試劑 Buffer C至裂解物, vortex thoroughly.
6. 添加 200μl of pre-chilled absolute ethanol, 攪拌均勻; precipitation may occur, but it will not affect subsequent experiments.
7. Transfer the obtained liquid into a DNA extraction and purification column (成套工具) (每次約650~700μl), 離心機在 >8,000 轉速為 1 分分鐘, 丟棄收集的廢液, and reinsert the collection tube into the DNA extraction and purification column (成套工具) 下一步.
8. Place the DNA extraction and purification column (成套工具) 進入收集管, 添加 300µl 洗滌緩衝液 1, 離心機在 >8,000 轉速為 1 分分鐘, 丟棄廢液, and reinsert the DNA extraction and purification column (成套工具) 進入管中進行下一步.

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

7. 添加 500µl 洗滌緩衝液 1to the DNA extraction and purification column (成套工具), 離心機在 14,000 轉速 (20,000×g) 為了 2 分分鐘, extend the centrifugation time appropriately to dry the membrane further.
8. Place the DNA extraction and purification column (成套工具) 放入新的離心管中, 打開蓋子, incubate at 65°C for 2 分分鐘; this step can be extended appropriately to evaporate ethanol as much as possible, preventing residual ethanol from affecting downstream experiments.
9. Elute 100µl 洗脫緩衝液到膜上, 離心機在 12,000 轉速為 2 分分鐘.

(筆記: 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, 再次離心 12,000 轉速為 2 minutes to increase DNA yield.)