[CD1021] SMOChem™ Deoxynucleotide (dNTP) Mix, 25 mM each (100 mM total), 500 µl x 6
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The SMOChem™ Deoxynucleotide (dNTP) Mix is an aqueous solution that contains an equimolar solution of ultrapure dATP, dCTP, dGTP and dTTP, each at a concentration of 25 mM at pH 8.5. The dNTP Mix is designed for many different molecular biology applications that involved in DNA synthesis or labeling, such as PCR, real-time PCR, DNA sequencing, reverse transcription, primer extension, and etc. The dNTP Mix is free of exo-deoxyribonuclease and endo-deoxyribonuclease as well as ribonuclease activity. The dNTP Mix offers the possibility to reduce the number of pipetting steps and the risk of reaction set up errors.
Detail
Description
The SMOChem™ Deoxynucleotide (dNTP) Mix is an aqueous solution that contains an equimolar solution of ultrapure dATP, dCTP, dGTP and dTTP, each at a concentration of 25 mM at pH 8.5. The dNTP Mix is designed for many different molecular biology applications that involved in DNA synthesis or labeling, such as PCR, real-time PCR, DNA sequencing, reverse transcription, primer extension, and etc. The dNTP Mix is free of exo-deoxyribonuclease and endo-deoxyribonuclease as well as ribonuclease activity. The dNTP Mix offers the possibility to reduce the number of pipetting steps and the risk of reaction set up errors.
Features
Ideal for PCR amplification and cDNA synthesis
Premixed solution
Nuclease and ribonuclease free
Applications
PCR amplification of DNA fragments
DNA fill-in reaction
DNA sequencing
Reverse transcription
One-step RT-PCR
Storage
-20°C for 36 months
Other Products
[DL2000] ExcelDye™ 6X DNA Loading Dye, Green, 5 ml x 2
Product Info
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Product Info
Description
The ExcelDye™ 6× DNA Loading Dye (Green) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains two dyes (Xylene cyanol FF and Orange G) for tracking the DNA migration. The Xylene cyanol FF and Orange G migrate at approximately 800 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.
Composition
0.03% Xylene cyanol FF
0.15% Orange G
10 mM Tris-HCl (pH 8.0)
60% glycerol
60 mM EDTA
Storage
4°C for 12 months -20°C for 36 months
Document
The ExcelDye™ 6× DNA Loading Dye (Green) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains two dyes (Xylene cyanol FF and Orange G) for tracking the DNA migration. The Xylene cyanol FF and Orange G migrate at approximately 800 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.
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.
Solid Phase Reversible Immobilization magnetic beads consist of paramagnetic particles coated with carboxyl groups that reversibly bind DNA. They are used for DNA purification because they are fast, simple and efficient. Our Magnetic Beads (PCR Purification) combines BioDynami’s proprietary chemistries with reversible DNA-binding properties of magnetic beads. The beads are developed for effective PCR fragment purification by removing primers and unwanted components such as salts, dNTPs, enzymes, and others.
Our Magnetic beads (PCR Purification) are optimized to selectively bind PCR fragments of 80 bp and larger and remove primers that are 30 nt and shorter. Purified PCR fragments are suitable for any downstream applications. The beads can be used for PCR cloning, PCR cleanup, or even PCR fragment concentration.
Features:
Effective purification of PCR fragments >80 bp
Removal of primers <30 nt
Removal of unwanted components
Flexibility: compatible with manual and automated processing
Applications:
Purification of PCR fragments
PCR cloning
Sequencing
Other applications requiring purified PCR fragments
Document
Solid Phase Reversible Immobilization magnetic beads consist of paramagnetic particles coated with carboxyl groups that reversibly bind DNA. They are used for DNA purification because they are fast, simple and efficient. Our Magnetic Beads (PCR Purification) combines BioDynami’s proprietary chemistries with reversible DNA-binding properties of magnetic beads. The beads are developed for effective PCR fragment purification by removing primers and unwanted components such as salts, dNTPs, enzymes, and others.
