Perkenalan
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. Atau, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidin-based lysis of samples and silica membrane–based purification of total RNA. Reagen Magzol, termasuk dalam kit, 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.
Detail
Spesifikasi
| Fitur | Spesifikasi |
| Fungsi utama | Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column |
| Aplikasi | RT-PCR, Northern Blot, poly A+purification, Perlindungan asam nukleat dan terjemahan in vitro |
| Metode pemurnian | Mini kolom putar |
| Teknologi pemurnian | Teknologi silika |
| Metode proses | petunjuk (sentrifugasi atau vakum) |
| Jenis sampel | Jaringan hewan, adherent cells, suspension cells, bakteri, dll. |
| Jumlah sampel | Eukaryotic culture cells: ≤ 10^7, Jaringan hewan:<100mg, Yeast culture cells:<5 x10^7, Bakteri:<10^9 |
| Volume elusi | ≥15μl |
| Waktu per lari | ≤40 minutes |
| Volume pembawa cairan per kolom | 800μl |
| Hasil pengikatan kolom | 100mg |
Prinsip
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. Akhirnya, 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. Akhirnya, the purified RNA was eluted by RNase free water.
Keuntungan
- Kualitas tinggi – one step Ekstraksi RNA reagent combined with silica gel column can obtain the highest concentration
- Cepat – beberapa sampel dapat diekstraksi 40 menit
- High applicability – samples including animals, tanaman, bakteri, sel, dll..
- Konsentrasi tinggi – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
Isi Paket
| Isi | R431002 | R431003 |
| Waktu Pemurnian | 50 Persiapan | 250 Persiapan |
| Kolom Mini RNA HiPure | 100 | 2 X 250 |
| 2ml Tabung Koleksi | 100 | 2 X 250 |
| Reagen Magzol | 60 ml | 270 ml |
| Penyangga RWC | 20 ml | 80 ml |
| Penyangga RW2* | 20 ml | 2 X 50 ml |
| Air Bebas RNase | 10 ml | 30 ml |
Penyimpanan dan Stabilitas
MagZol Reagent should be stored at 2-8°C upon arrival. Namun, penyimpanan jangka pendek (hingga 24 minggu) pada suhu kamar (15-25°C) tidak mempengaruhi kinerjanya. Komponen kit lainnya dapat disimpan pada suhu kamar (15-25°C) dan setidaknya stabil 18 bulan dalam kondisi ini.
Ulasan
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