SENO Sperm Sample 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.	SN0253	SN0254
DNA抽出カラム (セット)	50 (セット)	100 (セット)
Reagent Buffer SP	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
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 sperm sample ゲノムDNA抽出 kit provides a rapid and efficient purification solution for sperm sample DNA. It achieves sample DNA精製 effectively through a dedicated extraction buffer system combined with specific nucleic acid purification columns.

This kit can extract sperm sample DNA within 30 分, and the entire purification process does not 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.Reagent Buffer SP:This buffer should be stored long-term in an environment between 2℃ and 8℃

B.Reagent Buffer C 低温条件下で沈殿する可能性があります. It is recommended to heat at 65℃ for 5 分. After the precipitate dissolves, it can be used normally.

C.洗浄バッファー 1: ご使用の前に, add the specified amount of anhydrous ethanol as indicated on the reagent bottle label. Mark a check on the label to indicate the addition of anhydrous ethanol.

D. 溶出バッファーはaです 0.1X TEソリューション 最小限のEDTAを含む. EDTAがその後の実験に影響を与える場合, it is recommended to use sterile deionized water as a substitute for the elution buffer.

1. サンプル処理:

ピペット 200μl of frozen sperm, incubate at 75℃ for 10 minutes until the sperm sample is not viscous. で遠心分離します。 12000 の回転数 5 分, aspirate the supernatant as much as possible, and the sperm cells will sediment at the bottom of the tube.

2. Add to the sediment from the previous step: 400μl Reagent Buffer SP, 20μl Proteinase K (10 mg/ml), 20μLRNasea (10 mg/ml).Digest at 65℃ for 10 分, invert and mix 6-7 times during digestion until the sample is fully digested.
3. 等量を追加します Reagent Buffer C and an equal volume of anhydrous ethanol, and mix well by pipetting.

(例えば: If you add 400μl of Reagent Buffer IV, you will need to add approximately 460μl of Reagent Buffer C and 460μl of anhydrous ethanol. A small amount of precipitation may form after adding Reagent Buffer C, but it does not affect subsequent experiments.)

4. Transfer the obtained liquid to a DNA extraction purification column (cassette) (毎回約650〜700μl). Let it stand at room temperature for 2 分, オーバーでの遠心 8,000 の回転数 1 分, discard the collected waste liquid, and re-insert the collection tube into the purification column for the next step.
5. ステップを繰り返します 4, adding the remaining liquid to the DNA extraction purification column (cassette), オーバーでの遠心 8,000 の回転数 1 分, discard the waste liquid and the collection tube.
6. Place the DNA extraction purification column (cassette) into the collection tube, 追加 300 洗浄バッファーμl 1, オーバーでの遠心 8,000 の回転数 1 分, discard the waste liquid, and place the DNA extraction purification column (cassette) back into the tube for the next step.

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

7. 追加 500洗浄バッファーμl 1 to the DNA extraction purification column (cassette), 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 more thoroughly.
8. Place the DNA extraction purification column (cassette) 新しい遠心チューブに, open the lid, incubate at 65℃for 2 分. Extend this step if necessary to evaporate ethanol as much as possible, preventing ethanol residue from affecting downstream experiments.
9. Drop-suspend 50-100μL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. Using 50μl of Elution Buffer to elute DNA can increase DNA concentration but decreases the overall DNA yield. 2. The eluted DNA can be reapplied to the DNA extraction purification column, centrifuged at 12,000 の回転数 2 minutes again, to increase DNA yield.)