introduzione
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/guanidina-based lysis of samples and silica membrane–based purification of total RNA. Reagente MagZol, 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.
Dettagli
Specifiche
Caratteristiche | Specifiche |
Funzioni principali | Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column |
Applicazioni | RT-PCR, Northern Blot, purificazione poli A+, protezione degli acidi nucleici e traduzione in vitro |
Metodo di purificazione | Mini colonna rotante |
Tecnologia di purificazione | Tecnologia della silice |
Metodo di processo | Manuale (centrifugazione o vuoto) |
Tipo di campione | Animal tissues, adherent cells, suspension cells, batteri, eccetera |
Importo del campione | Eukaryotic culture cells: ≤ 10^7, Animal tissue:<100mg, Yeast culture cells:<5 x10^7, Bacteria:<10^9 |
Volume di eluizione | ≥15μl |
Tempo per corsa | ≤40 minutes |
Volume di trasporto del liquido per colonna | 800ml |
Resa legante della colonna | 100mg |
Principio
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. La membrana filtrante adsorbita con acido nucleico viene lavata per rimuovere la proteina residua e il sale. Finalmente, l'acido nucleico adsorbito sulla membrana del filtro può essere lavato via con un tampone a basso contenuto di sale (come il tampone TE) o acqua. L'acido nucleico ottenuto ha un'elevata purezza e può essere utilizzato direttamente in vari esperimenti a valle.
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. Finalmente, the purified RNA was eluted by RNase free water.
Vantaggi
- Alta qualità – one step Estrazione dell'RNA reagent combined with silica gel column can obtain the highest concentration
- Veloce – possono essere estratti diversi campioni 40 minuti
- High applicability – samples including animals, impianti, batteri, cellule, eccetera.
- High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
Contenuto del kit
Contenuti | R431002 | R431003 |
Tempi di purificazione | 50 Preparativi | 250 Preparativi |
HiPure RNA Mini Columns | 100 | 2 X 250 |
2ml Provette di raccolta | 100 | 2 X 250 |
Reagente MagZol | 60 ml | 270 ml |
Buffer RWC | 20 ml | 80 ml |
Buffer RW2* | 20 ml | 2 X 50 ml |
Acqua libera da RNasi | 10 ml | 30 ml |
Conservazione e stabilità
MagZol Reagent should be stored at 2-8°C upon arrival. Tuttavia, conservazione a breve termine (fino a 24 settimane) a temperatura ambiente (15-25°C) does not affect its performance. I restanti componenti del kit possono essere conservati a temperatura ambiente (15-25°C) e sono stabili almeno per 18 mesi in queste condizioni.
Recensioni
Non ci sono ancora recensioni.