The Primerdesign™ genesig® Kit for Enterocytozoon bieneusi (E. bieneusi) genomes is designed for the in vitro quantification of E. bieneusi genomes. The kit is designed to have the broadest detection profile possible whilst remaining specific to the E. bieneusi genome. The primers and probe sequences in this kit have 100% homology with a broad range of E. bieneusi sequences based on a comprehensive bioinformatics analysis.
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.
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Plasmid Purification Magnetic Beads (RNA Depletion)
Product Info
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Product Info
Plasmid Purification Magnetic Beads (RNA Depletion)
Plasmid isolation from bacterial cultures is one of the most popular techniques in biomedical research and pharmaceutical industries. However, it is common that the isolated plasmid DNA is usually contaminated with varied degrees of host RNA. Plasmid purification is necessary to reduce the impact on downstream applications by removing RNA contamination.
We have developed a simple reagent to completely remove RNA contamination in the isolated plasmid samples using Solid Phase Reversible Immobilization (SPRI) beads. SPRI beads consist of paramagnetic particles coated with carboxyl groups that reversibly bind DNA. Our Plasmid Purification Magnetic Beads (RNA Depletion) combines BioDynami’s proprietary chemistries with the reversible DNA-binding properties of SPRI magnetic beads. The reagent removes RNA and recovers the plasmid in the same step. Moreover, unwanted components such as salts, dNTPs, proteins, enzymes, and other impurities can also be removed simultaneously.
Plasmid can be used for downstream applications such as enzymatic digestion, transformation, transfection and molecular cloning etc. The beads can be an effective and inexpensive reagent for bacterial RNA depletion for routine plasmid purification.
Features
Effective depletion of bacterial RNA by RNase
High recovery rate of plasmid DNA by magnetic beads
Removal of unwanted components and impurities
Simple and fast beads-based protocol
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Plasmid isolation from bacterial cultures is one of the most popular techniques in biomedical research and pharmaceutical industries. However, it is common that the isolated plasmid DNA is usually contaminated with varied degrees of host RNA. Plasmid purification is necessary to reduce the impact on downstream applications by removing RNA contamination.
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.
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The APOPercentage™ Apoptosis kit is a dye-based, colorimetric assay for detection and measurement of apoptosis (programmed cell death) during in-vitro cell culture.
Propargyl-PEG2-NHS ester is a PEG linker with a propargyl group that can participate in copper catalyzed azide-alkyne Click Chemistry and NHS ester that can be used to label amine-containing entitites.
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Propargyl-PEG2-NHS ester is a PEG linker with a propargyl group that can participate in copper catalyzed azide-alkyne Click Chemistry and NHS ester that can be used to label amine-containing entitites.
