動物/細胞ゲノムDNAミニ抽出キット

1. 試薬キットのコンポーネント

仕様	50T	100T
猫. いいえ.	SN0221	SN0222
DNA抽出カラム (セット)	50 (セット)	100 (セット)
Reagent Buffer IV	20 ミリリットル	2×20 ミリリットル
Reagent Buffer Cplus	30 ミリリットル	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	2x1ml
RNaseA	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 Animal/Tissue DNA精製 Kit offers a rapid and effective solution for animal tissue and cell culture DNA purification, widely applicable across animal tissues and cell cultures.

This DNA rapid purification kit swiftly extracts total DNA from animals and cells, yielding DNA directly usable for PCR, サザンブロット, and similar applications. The purification process does not require toxic agents like phenol-chloroform, making this DNA purification kit highly suitable for various other samples.

5. 実験原則と手順

6. 抽出プロセス

Before Starting the Experiment:

あ. Reagent Buffer IV tends to precipitate under low-temperature conditions. It is recommended to heat at 65℃ for 5 分. After the precipitation dissolves, it can be used normally.

B. 使用する前, 指定された量の無水エタノールを追加します ウォッシュバッファ 1 as indicated on the reagent bottle label, 無水エタノールの追加を示すためにラベルのチェックをマークします.

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

1. サンプル処理:

あ. Take cell culture, で遠心分離する 12,000 の回転数 1 minute to collect cells, and aspirate the supernatant as much as possible. Add 250μl of Reagent Buffer IV and 10μl of RNaseA (10 mg/ml) to the collected cells, thoroughly suspend, and incubate at 65°C for 10 分.

B. Grind tissue not exceeding 25 mg into a fine powder using liquid nitrogen. Add 200μl of Reagent Buffer IV, 20μl of Proteinase K (10 mg/ml), and 10μl of RNaseA (10 mg/ml) to the powder, よく振る, and incubate at 65°C for 30 分, shaking the mixture four times during this period.

2. 追加 200μl of Reagent Buffer Cplus to the lysate and mix well. If a white precipitate appears, it can be left undisturbed; it won’t affect subsequent experiments once the precipitate disappears.

3. 追加 200無水エタノールのμL and mix thoroughly. Some precipitation might occur, but it won’t affect subsequent experiments.

4. Apply the obtained liquid to the DNA extraction purification column (sleeve) (approximately 650~700μl each time), let it sit at room temperature for 2 分, 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 purification column for the next step.

5. Place the DNA extraction purification column (sleeve) into a new collection sleeve, 追加 700μlの ウォッシュバッファ 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 place the DNA extraction purification column (sleeve) back into the sleeve for the next step.

(注記: Confirm the addition of anhydrous ethanol to Rinse Buffer 1.)

6. 追加 500μlの ウォッシュバッファ 1 to the DNA extraction purification column (sleeve), で遠心分離する 12,000 の回転数 30 秒, extend centrifugation time appropriately to ensure the membrane is drier.

(注記: Ethanol has a significant impact on subsequent experiments; したがって, 膜の乾燥が非常に重要です. 遠心分離後, ensure no ethanol remains before elution. Discard the waste and collection tubes afterward. After rinsing with Wash Buffer 1, the membrane on the DNA purification column should have a faint color. Carefully remove the DNA purification column after centrifugation, ensuring it doesn’t touch the collection tube to prevent ethanol contamination.)

7. Place the DNA purification column in a new centrifuge tube, add 100μl of Elution Buffer directly onto the membrane, で遠心分離する 12,000 の回転数 1 分, and collect the DNA.

(注記: Eluting DNA with 50μl of Elution Buffer can increase DNA concentration but decreases total DNA yield.)