This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.
Detail
Introduction
This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.
Details
Specifications
Features
Specifications
Main Functions
Extract Pathogen RNA/DNA from 0.5ml plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution for mNGS application, remove host background nucleic acid.
Applications
RT-PCR，Real Time PCR, biochip analysis, NGS
Products
Pathogen DNA / RNA
Purification method
Polydisperse magnetic beads
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Sample type
low/cell-free samples such as plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution
Sample amount
0.5 ml
Adaptive instrument
Nucleic acid extractor, pipetting workstation
Principle
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. 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 Buffer NFW.
Kit Contents
Contents
R667200B
R6672-02B
Purification Times
24 Preps
96 Preps
2ml Bead Tubes
24
96
Proteinase K
12 mg
50 mg
Protease Dissolve Buffer
1.8 ml
3 ml
Buffer SDS (20%)
1.8 ml
8 ml
MagBind Particles
0.6 ml
2.5 ml
Buffer MLB
15 ml
60 ml
Buffer MW1*
13 ml
44 ml
Buffer MW2*
6 ml
50 ml
Buffer AVE
5 ml
30 ml
Storage and Stability
MagBind Particles and Proteinase K Solution should be stored at 2–8°C upon arrival. However, short-term storage (up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for 18 months under these conditions.
Other Products
Stool Nucleic Acid Collection and Preservation Devices Dx
Product Info
Document
Product Info
Overview
CE-IVDR marked in accordance with the European Commission Regulation (EU) No. 2017/746.
Ideal for use in in vitro workflows
No need to immediately process samples
Nucleic acid preservation at room temperature over 2 years for DNA and 7 days for RNA
Ship stool samples at room temperature safely
Compatible with most DNA/RNA isolation methods
Norgen’s Stool Nucleic Acid Collection and Preservation Devices Dx are designed for the collection and preservation of nucleic acids from fresh stool specimens. The Stool Nucleic Acid Collection and Preservation Devices Dx – 50 contains 50 individual Stool Nucleic Acid Collection and Preservation Devices Dx. Each Stool Nucleic Acid Collection and Preservation Device Dx consists of Norgen’s Stool Nucleic Acid Collection and Preservation Tube Dx containing Norgen’s Stool Preservative in a liquid format. The user simply collects stool into the tubes (fill up to the line indicated on the tube) and mixes gently until the stool is well submerged under the preservative. The Stool Nucleic Acid Collection and Preservation Tube Dx is subsequently sent to the laboratory for DNA and/or RNA isolation and analysis. The stool DNA in preserved samples is stable for more than 2 years at room temperature, and the stool RNA in preserved samples is stable for 7 days at room temperature. DNA can be isolated from the preserved stool samples using Norgen’s Stool DNA Isolation Kit Dx (Cat. Dx27600) and RNA can be isolated from the preserved stool samples using Norgen’s Stool Total RNA Purification Kit Dx (Cat. Dx49500). These tubes are ideal for collecting and preserving DNA and RNA samples for in vitro diagnostic use for medical purposes.
NOTE: This product is not available for sale in the United States.
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.
How is apoptosis detected or measured?
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).
Biocolor’s APOPercentage assay is based on the latter. Further information can be found under the ‘Mode of Action’ Tab.
How does APOPercentage detect apoptosis?
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.
Cell membrane changes during apoptosis
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.
How are APOPercentage-labelled cells quantified?
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.
Limit of Detection
A single cell (via image analysis method)
Detection Method
Colorimetric (550nm) (Endpoint) or Image Analysis based
Measurements per kit
Sufficient for 4×24 well plates or 6×96 well plates
Suitable Samples
Adherent mammalian cells (in-vitro)
APOPercentage kit contents:
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.
Document
The APOPercentage™ Apoptosis kit is a dye-based, colorimetric assay for detection and measurement of apoptosis (programmed cell death) during in-vitro cell culture.
