How does the Agilent Bioanalyzer work?
The Agilent 2100 Bioanalyzer is a microfluidics-based platform for sizing, quantification and quality control of DNA and RNA. The fluorescent dye molecules intercalate into DNA or RNA strands. They are then detected by their fluorescence and translated into gel-like images (bands) and electropherograms (peaks).
What happens to nuclear DNA during apoptosis?
The degradation of cellular content is initiated in apoptotic cells and completed within engulfing cells. In apoptotic cells, caspase-mediated proteolysis cleaves protein substrates into fragments; nuclear DNA is partially degraded into nucleosomal units; and autophagy potentially contributes to apoptotic-cell removal.
How does RNA Bioanalyzer work?
How does the Bioanalyzer work? The bioanalyzer instrument detects biomolecules by laser-induced fluorescence. Each LabChip® contains an interconnected set of microchannels that is used for separation of nucleic acid fragments based on their size as they are driven through it electrophoretically.
What does a Bioanalyzer do RNA?
The Bioanalyzer is a chip-based capillary electrophoresis machine to analyse RNA, DNA, and protein. It is produced by Agilent and widely used, among other things, in RNA quality control measurements before downstream experiments like microarrays.
Which is activated for nuclear fragmentation in apoptosis?
Mechanism. The enzyme responsible for apoptotic DNA fragmentation is the Caspase-Activated DNase (CAD).
What are the stages of apoptosis?
To illustrate these apoptosis events and how to detect them, Bio-Rad has created a pathway which divides apoptosis into four stages: induction, early phase, mid phase and late phase (Figure 1).
What is in the apoptotic DNA ladder kit?
The Apoptotic DNA Ladder kit contains all of the reagents necessary for the preparation of cellular fractions for the production of apoptosis nucleosomal DNA ladders. The kit does not require the use of toxic phenol. The protocol involves: cell lysis; removal of cellular debris; and precipitation of nucleosomal DNA.
How to detect apoptosis using DNA ladder assay?
In our strategy, employing a practical DNA extraction protocol, DNA ladder assay could be applied as an easy/fast method for apoptosis detection. This improved method is able to detect apoptosis in a cost effective/timely manner without need for commercial kits and special equipment.
What is dnadna ladder assay?
DNA ladder assay is an easily available method and seems to be very useful for quick screening of apoptotic changes in cell populations. This method allows working with cell lysates and dose not require any special laboratory equipment. 11 It is a simple assay to perform, however being time consuming procedure,…
Can apoptosis be seen in gel electrophoresis by induction of apoptosis?
Apoptosis can be visualized as a ladder pattern of 180-200 bp due to DNA cleavage by the activation of a nuclear endonuclease by standard agarose gel electrophoresis.10,11Thus, we showed the formation of the DNA ladder in gel electrophoresis by induction of apoptosis in NIH-3T3 cell line (Fig. 1). Open in a separate window Fig. 1