The HiPure FastFilter Plasmid DNA Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS 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 a special 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. Up to 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 1mg plasmid DNA from 200ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR and labeling, etc. |
| Purification method | Maxi spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount | 100-200ml |
| Yield | 0.4-1mg |
| Elution volume | ≥500μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 20ml |
| Binding yield of column | 1mg |
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: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
Kit Contents
| Contents | P101402 | P101403 |
| Purification Times | 10 Preps | 50 Preps |
| RNase A | 20 mg | 40 mg |
| Buffer P1 | 140 ml | 2 x 350 ml |
| Buffer P2 | 140 ml | 2 x 350 ml |
| Buffer LEN3 | 70 ml | 350 ml |
| Buffer GBT | 120 ml | 550 ml |
| Buffer PW1 | 60 ml | 300 ml |
| Buffer PW2* | 50 ml | 4 x 100 ml |
| Elution Buffer | 20 ml | 120 ml |
| HiPure DNA Maxi Columns III | 10 | 50 |
| Lysate Clear Maxi Syringe | 10 | 50 |
| 50 ml Collection Tubes | 20 | 100 |
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these 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.
For any technical problems or customized products, please contact us.
F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here
The HiPure FastFilter Plasmid DNA Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS 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 a special 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. Up to 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).
Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.
This version of the kit is designed to detect and measure INSOLUBLE forms of collagen. Chose our Sircol 2.0 collagen kit if you need to analyse SOLUBLE collagen.
Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.
Sircol dye reagent contains Sirius Red – a linear anionic dye with sulphonic acid side chain groups. Under assay conditions the Sircol dye binds the basic groups of soluble collagen molecules. Maximal binding occurs in collagens possessing intact triple helix organisation as the highly ordered Gly-X-Yn helical structure of tropocollagen further contributes to dye binding. This results in a high degree of dye-collagen specificity. Affinity is progressively reduced during heat denaturation 4ºC due to the unwinding of the triple helix and formation of random chains.
Step 1. Samples being assayed for insoluble collagen must first undergo a 2-3 hour pre-treatment with Sircol Fragmentation reagent. This converts insoluble collagen into water-soluble gelatin can then be assayed.
Step 2. Addition of Sircol Dye Reagent to these pre-treated insoluble collagen samples results in the formation of a denatured collagen-dye complex. This complex then precipitates during the dye incubation period and is subsequently isolated by centrifugation, followed by washing to remove unbound dye. The Denatured collagen-bound dye is then eluted and measured spectrophotometrically.
Step 3. The insoluble collagen content of unknown samples is quantified by comparison against a calibration curve prepared using a the denatured collagen standard supplied with the kit.
Assay range
100 – 1000 µg/ml
100µg/ml
Colorimetric Detection (556nm) (Endpoint)
110 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).
The assay can be used to assess the rate of production of newly laid down collagen fibres during periods of rapid growth, development, tissue repair, remodeling and wound healing. Sources of material includes tissues, bone and calcified tissue.
*Insoluble collagens must be converted into soluble form prior to assay. Instructions and regents are provided with the kit., depending on sample this will require prior salt/acid/acid-pepsin extraction.
**non-mammalian collagens may result in a reduced limit of detection. We recommend use of an assay standard matched to the species under assay.
Many customers have found that the straightforward sample processing and analysis of Sircol make it a good alternative to conventional hydroxyproline analysis.
This kit is designed for research use only. Not for use in diagnostic procedures.
Kit requires access to a centrifuge, water bath / heated block, as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm.
Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
1. Sircol Dye Reagent (1x110ml)
2. Denatured Collagen Reference Standard (1x5ml, 1.0mg/ml)
3. Acid-Salt Wash Reagent (1x20ml)
4. Fragmentation Reagent (1x110ml)
5. Alkali Reagent (1x110ml)
6. 2ml screw-cap tubes for preparation of samples.
7. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
As collagens mature, they become increasingly crosslinked and insoluble – characteristics necessary for key biophysical role that collagen plays in living organisms. Biocolor’s Sircol™ INSOLUBLE Collagen Kit is a dye-binding assay designed for accurate quantification and measurement such collagens. It is ideal for analyzing crosslinked / insoluble collagens from sources such as tissues, bone, and calcified tissue.
Description
The ExcelTaq™ Blood Direct DNA Polymerase is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct DNA Polymerase is highly tolerant in the presence of PCR interfering/ inhibiting substances in blood, such as IgG, hemoglobin, and lactoferrin. ExcelTaq™ Blood Direct DNA Polymerase is compatible with most anticoagulants, such as citrate, EDTA, and heparin (Fig. 1). The ExcelTaq™ Blood Direct DNA Polymerase includes a pair of Positive Control Primers (CCR5) that are compatible with primate blood samples.
Features
Applications
Storage
-20°C for 24 months
4℃ for 6 months
The ExcelTaq™ Blood Direct DNA Polymerase is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct DNA Polymerase is highly tolerant in the presence of PCR interfering/ inhibiting substances in blood, such as IgG, hemoglobin, and lactoferrin. ExcelTaq™ Blood Direct DNA Polymerase is compatible with most anticoagulants, such as citrate, EDTA, and heparin (Fig. 1). The ExcelTaq™ Blood Direct DNA Polymerase includes a pair of Positive Control Primers (CCR5) that are compatible with primate blood samples.