Blood 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 Blood DNA純化 Reagent Kit offers a rapid and effective method for purifying DNA from blood and other bodily fluids. By lysing red blood cells with a red blood cell lysis buffer, DNA can be efficiently collected.

The Blood DNA Rapid Purification Kit can extract total DNA from whole blood (包括血液, 血清, 電漿, and other bodily fluids) 之內 30 分分鐘. The entire purification process doesn’t require toxic reagents such as phenol-chloroform. 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. Red Blood Cell Lysis Buffer 1x Preparation: Dilute the 10x Red Blood Cell Lysis Buffer to 1x volume using RNase-free water.

乙. Reagent Buffer B and Reagent Buffer C may precipitate at low temperatures. It is recommended to heat at 65℃ for 5 minutes to dissolve the precipitate before use.

C. Prior to using 洗滌緩衝液 1, add the specified amount of anhydrous ethanol as indicated on the reagent bottle label, and mark a check on the label to indicate ethanol addition.

D. 洗脫緩衝液是 0.1x TE 解決方案 with minimal EDTA. EDTA是否可能影響後續實驗, it is suggested to substitute Elution Buffer with sterile deionized water.

1. 樣品處理: (收集 0.1-5 ml blood samples)

A. 添加 2-3 times the volume of 1x Red Blood Cell Lysis Bufferto the blood or sample (Dilute the Red Blood Cell Lysis Buffer 10x to 1x before use), mix thoroughly, 離心機在 12,000 轉速為 1 分分鐘, carefully remove the supernatant. The precipitate should theoretically be white or pale red. Add Reagent Buffer B to the precipitate (for 0.1-1ml blood samples, 添加 200微升試劑緩衝液B; for 1-2ml blood samples, 添加 400微升 試劑緩衝液B).

乙. If handling samples from low-level organisms like poultry or birds with nucleated red blood cells, a smaller sample volume is needed. In such cases, omit the 1x Red Blood Cell Lysis Buffer and directly add 400微升of Reagent Buffer B and 20微升 of RNaseA (10 毫克/毫升), mix thoroughly, 並在室溫下孵育 5 分分鐘.

2. 添加 10微升RNaseA (10 毫克/毫升), 20微升 蛋白酶K (10 毫克/毫升), mix thoroughly, digest at 65℃ for 10 分分鐘. During this period, 反轉並混合 2-3 times until digestion is complete.

3. 添加 200微升of Reagent Buffer C 至裂解物, mix by pipetting, 添加 200微升 of anhydrous ethanol, mix by pipetting.

4. Apply the obtained liquid to the DNA extraction purification column (放) (每次約650~700μl), 離心機在 12,000 轉速為 30 秒, 丟棄收集的廢棄物, and re-insert the collection tube into the DNA extraction purification column (放) 下一步.

5. 添加 700微升of inhibition removal buffer, 離心機在 12,000 轉速為 30 秒, 丟棄廢棄物.

6. 放置DNA萃取純化管柱 (放) in a new collection tube, 添加 500微升of Wash Buffer 1, 離心機在 12,000 轉速為 30 秒, 丟棄廢棄物, 並重新插入DNA萃取純化管柱 (放) into the collection tube for the next step.

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

7. 重複步驟 6.

8. 放置DNA萃取純化管柱 (放) 放入新的離心管中, uncover, incubate at 65℃ in a water bath for 2 分分鐘. Extend this step appropriately to evaporate ethanol as much as possible to prevent ethanol residue from affecting downstream experiments.

9. Suspend 50-100微升of Elution Buffer onto the membrane of the column, 離心機在 12,000 轉速為 1 分分鐘, and collect the DNA.

(筆記: 1. Eluting DNA with 50微升 of Elution Buffer can increase DNA concentration but reduces total DNA yield; 2. The eluted DNA wash can be reapplied to the DNA extraction purification column. 離心機在 12,000 轉速為 1 minute again to increase DNA yield.