Introduction

The HiPure Fastfilter ADN plasmidique Midi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-FDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes aspecial filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Jusqu'à 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.

Détails

Caractéristiques

Principe

The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparationand clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. Après avoir lavé les protéines et autres impuretés, l'acide nucléique a finalement été élué avec un tampon à faible teneur en sel (10mmTris, pH9,0, 0.5mmEDTA).

Avantages

• Haute pureté – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc..
• Rapide – it takes only 15-60 minutes to complete the isolation
• Haut rendement – up to 70µg plasmid can be binded in one column

Contenu du kit

Stockage et stabilité

The kit components can be stored dry at room temperature (15-25°C) et sont stables pendant au moins 18 mois dans ces conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.

Guide d'achat