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/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, 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.
Specifications
| Features | Specifications |
| Main Functions | Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column |
| Applications | RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Animal tissues, adherent cells, suspension cells, bacteria, etc |
| Sample amount | Eukaryotic culture cells: ≤ 10^7, Animal tissue:<100mg, Yeast culture cells:<5 x10^7, Bacteria:<10^9 |
| Elution volume | ≥15μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
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. Finally, 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. Finally, the purified RNA was eluted by RNase free water.
Advantages
Kit Contents
| Contents | R431002 | R431003 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 100 | 2 x 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| MagZol Reagent | 60 ml | 270 ml |
| Buffer RWC | 20 ml | 80 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
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/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, 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.
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target
Accurate controls to confirm findings
150 reactions
For the determination of Escherichia coli in drinking water and its source water by multiple tube fermentation method.
MUG permits the rapid detection of Escherichia coli when the medium is observed for fluorescence using a UV light. MUG is hydrolyzed by the enzyme β-glucuronidase which is produced by E. coli to yield a fluorescent end product 4- methylumbelliferone. Trypticase provides the essential nutrients. Lactose is the fermentable carbohydrate. Sodium chloride maintains the osmotic equilibrium. The medium has a strong buffering system to control the pH in the presence of fermentative action. The bile salts inhibit gram-positive bacteria especially the Bacillus species and faecal Streptococci.
| Ingredients | /liter |
| Trypticase or tryptose | 20 g |
| Bile salts No. 3 | 1.5 g |
| Lactose | 5 g |
| K2HPO4 | 4 g |
| KH2PO4 | 1.5 g |
| NaCl | 5 g |
| 4-methylumbelliferyl-β-D-glucuronide (MUG) | 50mg |
| pH6.9±0.2 at 25°C | |
Weigh 37g of dry powder of this product, add 1L of distilled water or deionized water, stir, heat and boil until
completely dissolved, sterilize at 115℃ for 20min, cool to room temperature and set aside.
The following quality control strains were inoculated and cultured at 44.5℃±0.5℃ for 24h. The results are as follows:
| Quality control strains | Growth |
| Escherichia coli ATCC25922 | Blue-white fluorescence is produced under 366nm UV light |
| Salmonella typhimurium ATCC14028 | No fluorescence under 366nm UV light |
| Enterococcus faecalis ATCC29212 | Clear, no fluorescence |
2-30℃，Keep container tightly closed, avoid direct sunlight.
Use before expiry date on the label.
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
Intended Use For the determination of Escherichia coli in drinking water and its source water by multiple tube fermentation method. Principle and Interpretation MUG permits the rapid detec……