導入
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/グアニジン-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.
詳細
仕様
特徴 | 仕様 |
主な機能 | 分離miRNAおよびその他の小さなRNA分子(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column |
アプリケーション | RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation |
精製方法 | ミニ スピンカラム |
精製技術 | シリカ技術 |
加工方法 | マニュアル (遠心分離または真空) |
サンプルの種類 | 動物組織, adherent cells, suspension cells, 細菌, 等 |
サンプル量 | Eukaryotic culture cells: ≤ 10^7, 動物組織:<100mg, Yeast culture cells:<5 x10^7, 細菌:<10^9 |
溶出量 | ≥15μl |
実行あたりの時間 | ≤40 minutes |
カラムあたりの液体搬送量 | 800μl |
カラムの結合収量 | 100μg |
原理
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such asguanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein andother impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. ついに, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
This kit combines acid/guanidine (MagZol) extraction technology with glass fiber filter membrane purification, which can improve the extraction effect of complex samples and samples with low RNA content. After the sample is treated with MagZol reagent and chloroform, the supernatant is added with ethanol to provide appropriate binding conditions, then transferred to the purification column and centrifuged. Macromolecular RNA can be efficiently bound to the membrane. Collect the filtrate containing small RNA, add more ethanol to adjust the binding capacity of small RNA, the pollutants can be efficiently washed away by second cleaning. ついに, the purified RNA was eluted by RNase free water.
利点
- 高品質 – one step RNA抽出 reagent combined with silica gel column can obtain the highest concentration
- 速い – いくつかのサンプルを抽出できます 40 分
- High applicability – samples including animals, 植物, 細菌, 細胞, 等.
- 高濃度 – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
キット内容
コンテンツ | R431002 | R431003 |
浄化時間 | 50 準備 | 250 準備 |
Hipure RNAミニ柱 | 100 | 2 バツ 250 |
2ml 採取管 | 100 | 2 バツ 250 |
MagZol Reagent | 60 ミリリットル | 270 ミリリットル |
Buffer RWC | 20 ミリリットル | 80 ミリリットル |
バッファRW2* | 20 ミリリットル | 2 バツ 50 ミリリットル |
RNaseフリーウォーター | 10 ミリリットル | 30 ミリリットル |
保管と安定性
MagZol Reagent should be stored at 2-8°C upon arrival. しかし, 短期保管 (まで 24 週) 室温で (15-25℃) パフォーマンスには影響しません. 残りのキットコンポーネントは室温で保存できます (15-25℃) 少なくとも一定期間は安定しています 18 この状況下で数か月.
レビュー
まだレビューはありません.