Perkenalan
- Cepat, Sederhana, and Cost-Effective: Offers a rapid, easy-to-use, and economical method for DNA plasmid 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, menghemat waktu dan tenaga.
- No Phenol Extraction or Ethanol Precipitation: Bypasses the need for phenol extraction and ethanol precipitation, simplifying the workflow and reducing hands-on time.
- DNA berkualitas tinggi: Yields high-quality plasmid DNA eluted in a small volume of Tris buffer or water, ensuring optimal performance in downstream applications.
Spesifikasi
Fitur | Spesifikasi |
Fungsi utama | Isolasi hingga 35μg DNA plasmid dari 1-5ml kultur bakteri |
Aplikasi | Pencernaan enzim, pengurutan, PCR, kloning, dll.. |
Metode pemurnian | Mini kolom putar |
Teknologi pemurnian | Teknologi silika |
Metode proses | petunjuk (sentrifugasi atau vakum) |
Jenis sampel | Plasmid konvensional, plasmid kurang dari 30KB |
Jumlah sampel | Plasmid salinan tinggi: 1-5ml culture medium Low copy number plasmid: 5-10ml media kultur |
Menghasilkan | 5-35mg |
Volume elusi | ≥30μl |
Waktu per lari | Menyelesaikan 1-24 sampel di 30 menit |
Volume pembawa cairan per kolom | 800μl |
Hasil pengikatan kolom | 35mg |
Prinsip
- 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, Dan (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.
Keuntungan
- Kemurnian tinggi – Plasmid yang dimurnikan dapat langsung digunakan dalam pengurutan, PCR pencernaan enzim, dll..
- Cepat – hanya dibutuhkan 1 menit untuk mendapatkan supernatan dengan larutan yang dioptimalkan (10 menit untuk merek lain)
- Hasil tinggi – hingga 35µg plasmid dapat diikat dalam satu kolom
Isi Paket
Isi | P100102 | P100103 |
Waktu Pemurnian | 100 Persiapan | 250 Persiapan |
RNase A | 5 mg | 10 mg |
Penyangga P1 | 30 ml | 80 ml |
Penyangga P2 | 30 ml | 80 ml |
Penyangga P3 | 40 ml | 100 ml |
Penyangga PW1 | 60 ml | 140 ml |
Penyangga PW2* | 20 ml | 50 ml |
Penyangga Elusi | 15 ml | 30 ml |
Kolom Mini DNA HiPure II | 100 | 250 |
2 ml Tabung Koleksi | 100 | 250 |
Penyimpanan dan Stabilitas
- Kondisi penyimpanan: Kit components can be stored dry at room temperature (15-25°C) and remain stable for a minimum of 18 bulan dalam kondisi ini.
- Buffer Precipitates: In case of any precipitates forming in the buffers, warm them at 37°C to dissolve effectively.
- Buffer P1 Stability: Setelah penambahan RNase A, Buffer P1 remains stable for up to 6 bulan bila disimpan pada suhu 2-8°C.
Data Eksperimen
Panduan Pembelian
Nama | TIDAK KUCING | Jumlah sampel | Jenis kolom | Volume elusi | Menghasilkan | Waktu per lari | Persyaratan sentrifugasi |
Kit Mini Plasmid HiPure | P1001 | 1-5ml | 1.5kolom ml | 30-50μl | 5–35µg/5ml | ≤30 menit | mesin sentrifugal kecil |
Kit Mini Plasmid HiPure II | P1002 | 5-15ml | 1.5kolom ml | 75-100μl | 20-80mg/15ml | ≤30 menit | mesin sentrifugal kecil |
HiPure Plasmid Plus 96 Perlengkapan | P1006 | 1-1.5ml | 96 piring sumur | 70-150μl | 1-15μg/1ml | ≤60 menit | 96-centrifuge pelat sumur |
Kit Midi Plasmid Fastfilter HiPure | P1013 | 30-50ml | 15kolom ml | 0.3-0.8ml | 50–250µg/50ml | ≤60 menit | 15ml sentrifugasi |
Kit Maxi Plasmid Fastfilter HiPure | P1014 | 100-200ml | 50kolom ml | 0.5-1.5ml | 0.4–1mg/200ml | ≤60 menit | 50ml sentrifugasi |
Ulasan
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