血液ゲノムDNA抽出キット

1. 試薬キットのコンポーネント

仕様	50T	100T
猫. いいえ.	SN0219	SN0220
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 × 30ml
10×Red Blood Cell Lysis Buffer	60 ミリリットル	2 × 60ml
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	2x1ml
RNaseA	1ミリリットル	2x1ml
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 Blood DNA精製 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 (血液を含む, 血清, プラズマ, and other bodily fluids) 内で 30 分. The entire purification process doesn’t require toxic reagents such as phenol-chloroform. 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. Red Blood Cell Lysis Buffer 1x Preparation: Dilute the 10x Red Blood Cell Lysis Buffer to 1x volume using RNase-free water.

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

1. サンプル処理: (集める 0.1-5 ml blood samples)

あ. 追加 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), 十分に混ぜる, で遠心分離する 12,000 の回転数 1 分, 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, 追加 200μLReagent Buffer B; for 1-2ml blood samples, 追加 400μL Reagent Buffer B).

B. 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 400μLof Reagent Buffer B and 20μL of RNaseA (10 mg/ml), 十分に混ぜる, and incubate at room temperature for 5 分.

2. 追加 10μLRNaseA (10 mg/ml), 20μL プロテイナーゼK (10 mg/ml), 十分に混ぜる, digest at 65℃ for 10 分. During this period, invert and mix 2-3 times until digestion is complete.

3. 追加 200μLof Reagent Buffer C to the lysate, ピペッティングでミックスします, 追加 200μL of anhydrous ethanol, ピペッティングでミックスします.

4. Apply the obtained liquid to the DNA extraction purification column (セット) (approximately 650~700μl each time), で遠心分離する 12,000 の回転数 30 秒, 収集された廃棄物を廃棄します, and re-insert the collection tube into the DNA extraction purification column (セット) for the next step.

5. 追加 700μLof inhibition removal buffer, で遠心分離する 12,000 の回転数 30 秒, 廃棄物を捨てます.

6. Place the DNA extraction purification column (セット) in a new collection tube, 追加 500μLof Wash Buffer 1, で遠心分離する 12,000 の回転数 30 秒, 廃棄物を捨てます, and re-insert the DNA extraction purification column (セット) into the collection tube for the next step.

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

7. ステップを繰り返します 6.

8. Place the DNA extraction purification column (セット) 新しい遠心チューブに, uncover, incubate at 65℃ in a water bath for 2 分. Extend this step appropriately to evaporate ethanol as much as possible to prevent ethanol residue from affecting downstream experiments.

9. Suspend 50-100μLof Elution Buffer onto the membrane of the column, で遠心分離する 12,000 の回転数 1 分, and collect the DNA.

(注記: 1. DNAを溶出します 50μL 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. で遠心分離します。 12,000 の回転数 1 minute again to increase DNA yield.