This product is suitable for extracting total viral nucleic acid from cell-free/low-content cell biological samples such as body fluids, serums, plasma, soaking solutions, tissue homogenate supernatant, and culture supernatant. The Purified DNA/RNA is used for RT-PCR and PCR detection.
Specifications
| Features | Specifications |
| Main Functions | IVD5412 precast kit for kingfisher Flex |
| Applications | RT-PCR，PCR，NGS |
| Products | Viral DNA / RNA, body cell DNA / RNA |
| Purification method | Polydisperse magnetic beads |
| Purification technology | Magnetic beads technolog |
| Process method | Manual or automatic |
| Sample type | |
| Sample amount | 200μl |
| Adaptive instrument | Nucleic acid extractor, pipetting workstation |
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA/RNA is released into the lysate. After adding magnetic particles and binding solution, DNA/RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by Nuclease Free Water.
Advantages
Kit Contents
| Cat.No | Reagent | IVD5412-F-96 |
| PK/Carrier RNA | 1x 24 mg/Bottle | |
| Protease Dissolve Buffer Blue | 1.8 ml/Bottle | |
| Tip | 1 | |
| Sample Plate (DW Plate) | 500µl Buffer MLB/Well | 1 |
| Wash 1 Plate (DW Plate) | 500µl Buffer MW1/Well | 1 |
| Wash 2 Plate (DW Plate) | 500µl Buffer CW/Well | 1 |
| Beads Plate (DW Plate) | 500µl CW & 20µl MPN/Well | 1 |
| Elute Plate (KF Plate) | 90µl NFW/Well | 1 |
Storage and Stability
This kit is shipped and stored at room temperature and is valid for 12 months.
This product is suitable for extracting total viral nucleic acid from cell-free/low-content cell biological samples such as body fluids, serums, plasma, soaking solutions, tissue homogenate supernatant, and culture supernatant. The Purified DNA/RNA is used for RT-PCR and PCR detection.
These kits allow for the isolation of DNA from the blood of various species, including humans and will recover genomic DNA, mitochondrial DNA, viral DNA and bacterial DNA. The purified DNA is of excellent quality and yield and completely compatible with any downstream application including PCR, qPCR and genotyping.
Blood DNA Isolation Mini Kit
Norgen’s Blood DNA Isolation Mini Kit is designed for the rapid preparation of DNA from up to 200 µL of whole blood using a rapid spin column protocol. Preparation time for a single sample is less than 30 minutes and each kit contains sufficient materials for 50 preparations.
Blood DNA Isolation Midi Kit
Norgen’s Blood DNA Isolation Kit is designed for the rapid spin column preparation of DNA from 0.3 to 2 mL volumes of whole blood. Preparation time for a single sample is less than 30 minutes, and each kit contains sufficient materials for 20 preparations.
Blood DNA Isolation Maxi Kit
This kit is designed for the rapid preparation of DNA from 3 mL up to 10 mL of whole blood. Preparation time for a single sample is less than 30 minutes.
Blood DNA Isolation 96-Well Kit (High Throughput)
This kit provides a rapid method for the high-throughput isolation of DNA from up to 200 µL of whole blood. Fast and easy processing using either a vacuum manifold or centrifugation.
Blood DNA Isolation Kit (Magnetic Bead System)
Norgen’s Blood DNA Isolation Kit (Magnetic Bead System) is designed for the rapid preparation of genomic DNA from up to 200 µL of whole blood from various species, including human. Purification is based on magnetic beads as the separation matrix. Norgen’s magnetic beads bind DNA under optimized salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. The genomic DNA is preferentially purified from other cellular proteinaceous components. Typical yields of genomic DNA will vary depending on the cell density of the blood sample. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with downstream applications including real-time PCR, NGS and microarray analysis.
Blood DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)
96-well format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
Figure 1 / 19
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| Kit Specifications | |
| Maximum Blood Input | 200 µL |
| Average Yield (200 µL blood)* | 4-12 µg |
| Average Purity (OD260/280) | 1.8 – 1.9 |
| Time to Complete Purifications | 40 minutes (hands-on time Cat. 59800) 50 minutes (hands-on time Cat. 62600) |
* Average DNA yield will vary depending on the donor
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 1 year after the date of shipment when stored at room temperature.
| Component | Cat. 46300 (50 preps) | Cat. 46380 (100 preps) | Cat. 51400 (20 preps) | Cat. 31200 (12 preps) | Cat. 46350 (192 preps) | Cat. 59800 (50 preps) | Cat. 62600 (192 preps) |
|---|---|---|---|---|---|---|---|
| Lysis Buffer B | 20 mL | 2 x 20 mL | 2 x 40 mL | 2 x 110 mL | 2 x 40 mL | 20 mL | 1 x 40 mL 1 x 20 mL |
| Solution WN | 18 mL | 2 x 18 mL | 55 mL | 55 mL | 55 mL | 18 mL | 55 mL |
| Wash Solution A | 18 mL | 2 x 18 mL | 2 x 38 mL | 2 x 38 mL | 2 x 38 mL | – | – |
| Elution Buffer B | 30 mL | 2 x 30 mL | 30 mL | 30 mL | 2 x 30 mL | 15 mL | 1 x 30 mL 1 x 15 mL |
| Proteinase K in Storage Buffer | 1.2 mL | 2 x 1.2 mL | 4.4 mL | 8 mL | 4.5 mL | 1.2 mL | 4 mL |
| Magnetic Bead Suspension | – | – | – | – | – | 2 x 1.1 mL | 8.5 mL |
| Maxi Spin Columns | – | – | – | 12 | – | – | – |
| Mini Spin Columns | 50 | 100 | – | – | – | – | – |
| Midi Spin Columns | – | – | 20 | – | – | – | – |
| 96-Well Plate | – | – | – | – | 2 | – | 2 |
| Adhesive Tape | – | – | – | – | 4 | – | 2 |
| Collection Tubes | 50 | 100 | 20 | 12 | – | – | – |
| 96-Well Collection Plate | – | – | – | – | 2 | – | – |
| Elution Tubes | 50 | 100 | 20 | 12 | – | 50 | – |
| 96-Well Elution Plate | – | – | – | – | 2 | – | 2 |
| Product Insert | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
Apoptosis is an essentially normal physiological process that removes now redundant, cells, particularly during embryonic development and early growth. In adult animals the process removes cells that are irreparable. The apoptotic process is also involved in many major diseases such as cancer, where transformed tumour cells have their apoptotic process disabled, permitting cell cycling to continue unchecked. In contrast some forms of senile dementia may result from excessive apoptotic induction of neural cells.
The apoptotic process in mammalian cells is a rapid event (2‐4 hours). Within this short time span an apparently viable cell can be quietly dismantled, to disappear leaving no visible trace of its former existence.
An apoptosis cascade of activators, effectors and regulators has been identified. This in turn led to a range of apoptosis assays being devised to detect and monitor these events. Some laboratories will employ two distinct assays, one selected to detect early (initiation) apoptotic events, while a second assay will target a later (execution) event. Apoptosis assays, based on methodology, can be classified into four major inter‐linked groups:
[1] DNA fragmentation (electrophoresis and nick end labelling, TUNEL).
[2] Apoptotic proteases (fluorescently labelled antibodies to the caspases).
[3] Flow cytometric analysis (FACS, incorporating other group assays).
[4] Membrane alterations (phosphatidylserine flip).
Biocolor’s APOPercentage assay is based on the latter. Further information can be found under the ‘Mode of Action’ Tab.
The mammalian cell membrane has been described as a semi‐fluid mosaic structure, composed of phospholipids with a diverse group of inserted proteins and some cholesterol. The phospholipids are the major components of the membrane and are arranged in the form of a ‘bi‐layer’; which is asymmetric in composition, structure, and function.
To ensure normal transmembrane functions the phospholipids must be maintained in an asymmetric composition. The process is regulated by ‘flippases’, which catalyse the active transport of aminophospholipids from the outer to inner monolayer. However, in cells undergoing apoptosis, flippase is overwhelmed by the action of another enzyme, termed ‘floppase’ or ‘scramblase’. The net effect is a scrambling of the phospholipid distribution between the inner and outer monolayers.
The APOPercentage assay utilises an intense, pink-coloured dye reagent which is taken up during in-vitro culture by apoptosis-committed cells. This uptake occurs at the stage of Phosphatidylserine transmembrane movement, as produced by the flipflop mechanism. Dye uptake continues until blebbing occurs. No further dye can then enter the now defunct cell and the dye that has accumulated within the cell is not released (unlike necrotic cells which release dye).
Since the dye reagent is excluded or not retained by healthy or necrotic cells it therefore acts as a specific label for apoptotic cells.
Labelled apoptosis cells may then by conveniently analysed by the following methods:
Direct Analysis
The intense pink colour of the labelled cells can be visually assessed using brightfield microscopy. Apoptosis in substrate-adherent cell populations is therefore readily quantified using image analysis techniques. This technique is the most sensitive with the ability of detecting one single apoptotic cell per well.
Colorimetry protocol
Dye that accumulates within apoptotic cells is released into solution via addition of Dye Release Reagent. The concentration of this intracellular dye is then measured at 550nm using a microplate colorimeter/spectrophotometer.
NB: The APOPercentage assay kit does NOT require the use of a Flow Cytometer.
A single cell (via image analysis method)
Colorimetric (550nm) (Endpoint) or Image Analysis based
Sufficient for 4×24 well plates or 6×96 well plates
Adherent mammalian cells (in-vitro)
1. APOPercentage Dye (1x5ml)
2. Dye Release Reagent (1x150ml)
3. Phosphate Buffered Saline (PBS) (1x120ml)
4. 24-well starter plate.
5. Assay kit manual.
The Colorimetric Protocol requires a Microplate Colorimeter / Spectrophotometer.
Additional 96-well plates will be required for use when reading dye absorbance values.
The Direct Detection Protocol Requires an inverted stage microscope with an attached digital camera.
NB: Additional reagents (typically culture medium and suitable apoptosis treatments) may be required for sample preparation prior to assay. Consult manual or contact us for further details.
The APOPercentage™ Apoptosis kit is a dye-based, colorimetric assay for detection and measurement of apoptosis (programmed cell death) during in-vitro cell culture.