Uvod
- Brzo, Simple, and Cost-Effective: Offers a rapid, easy-to-use, and economical method for plasmid DNA miniprep suitable for routine molecular biology applications.
- Silica Membrane Technology: Utilizes silica membrane technology, eliminating the need for cumbersome steps associated with loose resins or slurries, streamlining the purification process.
- Immediate Usability: Purified plasmid DNA is immediately ready for downstream applications, saving time and effort.
- No Phenol Extraction or Ethanol Precipitation: Bypasses the need for phenol extraction and ethanol precipitation, simplifying the workflow and reducing hands-on time.
- High-Quality DNA: Yields high-quality plasmid DNA eluted in a small volume of Tris buffer or water, ensuring optimal performance in downstream applications.
Tehnički podaci
Značajke | Tehnički podaci |
Glavne funkcije | Isolation of up to 35μg plasmid DNA from 1-5ml bacterial culture |
Prijave | Enzyme digestion, sequencing, PCR, cloning, itd. |
Metoda pročišćavanja | Mini spin column |
Tehnologija pročišćavanja | Silika tehnologija |
Procesna metoda | Priručnik (centrifugiranje ili vakuum) |
Vrsta uzorka | Konvencionalni plazmid, plazmid manji od 30KB |
Količina uzorka | High copy plasmid: 1-5ml culture medium Low copy number plasmid: 5-10ml culture medium |
Prinos | 5-35µg |
Volumen eluacije | ≥30μl |
Vrijeme po vožnji | Complete 1-24 samples in 30 minuta |
Nosivi volumen tekućine po stupcu | 800µl |
Prinos vezivanja kolone | 35µg |
Načelo
- Alkaline Lysis: Begins with the alkaline lysis of bacterial cells, which breaks down the cell membrane and releases plasmid DNA into the lysate.
- Silica-Based DNA Binding: The released DNA is adsorbed onto the silica membrane in the presence of high salt, facilitating efficient binding of DNA to the membrane surface.
- Three Basic Steps: The procedure involves three main steps: (1) Preparation and clearing of the bacterial lysate using an alkaline method, (2) Transfer of the cleared lysate to a column for DNA binding, i (3) Washing to remove proteins and other impurities.
- Silica Membrane: Utilizes a unique silica membrane in the kit, which completely replaces traditional glass or silica slurries typically used in plasmid DNA minipreps.
- Elution with Low-Salt Buffer: After washing to remove impurities, nucleic acid, including plasmid DNA, is eluted from the silica membrane using a low-salt buffer containing 10mM Tris, pH 9.0, and 0.5mM EDTA.
Prednosti
- Visoka čistoća – pročišćeni plazmid može se izravno koristiti u sekvenciranju, enzyme digestion PCR, itd.
- Brzo – it takes only 1 minute to obtain supernatant by optimized solution (10 minutes for other brands)
- Visok prinos – up to 35µg plasmid can be bound in one column
Sadržaj kompleta
Sadržaj | P100102 | P100103 |
Vremena pročišćavanja | 100 Pripreme | 250 Pripreme |
RNaza A | 5 mg | 10 mg |
Buffer P1 | 30 ml | 80 ml |
Buffer P2 | 30 ml | 80 ml |
Buffer P3 | 40 ml | 100 ml |
Buffer PW1 | 60 ml | 140 ml |
Međuspremnik PW2* | 20 ml | 50 ml |
Elucijski pufer | 15 ml | 30 ml |
HiPure DNA mini kolone II | 100 | 250 |
2 ml cijevi za prikupljanje | 100 | 250 |
Skladištenje i stabilnost
- Storage Conditions: Kit components can be stored dry at room temperature (15-25°C) and remain stable for a minimum of 18 mjeseci pod ovim uvjetima.
- Buffer Precipitates: In case of any precipitates forming in the buffers, warm them at 37°C to dissolve effectively.
- Buffer P1 Stability: After the addition of RNase A, Buffer P1 remains stable for up to 6 months when stored at 2-8°C.
Experiment Data
Vodič za kupnju
Ime | MAČKA BR | Količina uzorka | Vrsta stupca | Volumen eluacije | Prinos | Vrijeme po vožnji | Zahtjevi za centrifugu |
HiPure Plasmid Mini Kit | P1001 | 1-5ml | 1.5ml stupca | 30-50μl | 5–35µg/5ml | ≤30 minuta | mala centrifuga |
HiPure Plasmid Mini Kit II | P1002 | 5-15ml | 1.5ml stupca | 75-100μl | 20-80µg/15ml | ≤30 minuta | mala centrifuga |
HiPure Plasmid Plus 96 Kit | P1006 | 1-1.5ml | 96 bunar ploča | 70-150μl | 1-15μg/1ml | ≤60 minuta | 96-well plate centrifuge |
HiPure Fastfilter Plasmid Midi Kit | P1013 | 30-50ml | 15ml stupca | 0.3-0.8ml | 50–250µg/50ml | ≤60 minuta | 15ml centrifuga |
HiPure Fastfilter Plasmid Maxi Kit | P1014 | 100-200ml | 50ml stupca | 0.5-1.5ml | 0.4–1mg/200ml | ≤60 minuta | 50ml centrifuga |
Recenzije
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