ArcticZymes RNA to DNA Ligase (ArcticZymes R2D Ligase™) is the first ligase on the market that is able to ligate DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the joinable ends. With its unique substrate specificity, ArcticZymes R2D Ligase allows the development of new technologies in molecular biology research, diagnostics, and manufacturing.
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ArcticZymes RNA to DNA Ligase (ArcticZymes R2D Ligase™) is the first ligase on the market that is able to ligate DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the joinable ends. With its unique substrate specificity, ArcticZymes R2D Ligase allows the development of new technologies in molecular biology research, diagnostics, and manufacturing.
Key Features:
Novel substrate specificity
RNA to DNA ligation
High purity
Detergent free
Efficient ligation of DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the ligatable ends of DNA and RNA
ATP dependent dsDNA ligase
Utilizes Mg2+ or Mn2+
Suggested Applications:
RNA capturing
In vitro transcription
NGS library prep
Transcriptome amplification
miRNA detection/analysis
RNA splint ligation
Other Products
[DM1100] ExcelBand™ 50 bp DNA Ladder, 500 μl
Product Info
Document
Product Info
Description
The DM1100 ExcelBand™ 50 bp DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA ladder DM1100 is composed of 17 individual DNA fragments: 1.5k, 1.2k, 1k, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, and 50 bp derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains two enhanced bands (500 bp and 200 bp) for easier reference. In addition, the low range Orange G tracking dye mimics the migration of a 50 bp dsDNA during electrophoresis is also added for real time monitoring.
Features
Sharp bands
Quick reference— enhanced bands
Ready-to-use— premixed with loading dye for direct loading
Stable— room temperature storage over 6 months
Source
Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA.
Range
50 ~ 1,500 bp
Concentration
54 µg/ 500 µl
Recommended loading volume
5 µl/ well
Storage
Room temperature for 6 months 4°C for 12 months -20°C for 36 months
Document
The DM1100 ExcelBand™ 50 bp DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA ladder DM1100 is composed of 17 individual DNA fragments: 1.5k, 1.2k, 1k, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, and 50 bp derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains two enhanced bands (500 bp and 200 bp) for easier reference. In addition, the low range Orange G tracking dye mimics the migration of a 50 bp dsDNA during electrophoresis is also added for real time monitoring.
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.
Opentrons Flex 8-Channel pipettes enable highly accurate pipetting, optimized for automation.
Flex pipettes use air displacement technology to offer highly accurate pipetting with pipette volume ranges from 1 to 50 µL and 5 to 1000 μL. Smart sensors support automatic calibration, real-time positioning, and error detection. Choose the pipettes that fit your workflow — the gantry supports any two Flex 1-Channel or 8-Channel Pipettes — and swap them out yourself when your workflow changes.
Document
Opentrons Flex 8-Channel pipettes enable highly accurate pipetting, optimized for automation.
Flex pipettes use air displacement technology to offer highly accurate pipetting with pipette volume ranges from 1 to 50 µL and 5 to 1000 μL. Smart sensors support automatic calibration, real-time positioning, and error detection. Choose the pipettes that fit your workflow — the gantry supports any two Flex 1-Channel or 8-Channel Pipettes — and swap them out yourself when your workflow changes.
