What Is Capillary Electrophoresis?
Summary
TLDRCapillary electrophoresis is a high-speed, high-throughput technique for separating DNA fragments using thin, polymer-filled capillaries. It allows for single-base resolution with minimal sample size. The process involves loading fluorescently labeled DNA into a capillary array, applying an electric field to move the fragments, and then detecting them as they pass a laser, with the fluorescence captured by a CCD camera. This method, ideal for STR analysis and Sanger sequencing, is showcased in the video by the Spectrum Compact CE system, emphasizing its efficiency and utility in life science and forensic labs.
Takeaways
- 🌟 Capillary electrophoresis is a technique for separating and identifying fluorescently labeled DNA fragments.
- 🧪 It uses thin polymer-filled capillaries instead of slab gels, allowing for stronger electric fields and faster separations.
- 🔋 The process offers high throughput and single base resolution, requiring only small sample quantities.
- 🔬 Applications include STR analysis, Sanger sequencing, and various other uses in life science and forensic labs.
- 🚀 The method begins with preparing and loading fluorescently labeled DNA samples into a capillary array.
- 🌐 An electric current is formed when the capillary array enters the sample wells, moving DNA fragments electrokinetically.
- 🔌 The ends of the capillary array are inserted into cathode and anode buffers, with a specific voltage and temperature applied for controlled migration.
- 📊 Fragments are separated by size, with the smallest moving the fastest, allowing for size-based analysis.
- 🔦 Detection occurs as fragments pass a detection window, where a laser excites the fluorescent dyes, and the emitted light is recorded.
- 📷 A CCD camera captures the fluorescence signature and migration time of each fragment, providing raw data for analysis.
- 💻 The data is sent to collection software as peaks, ready for analysis with various software programs.
- 🛠️ The animation is presented by Spectrum Compact, promoting a compact and powerful CE instrument for laboratory use.
Q & A
What is capillary electrophoresis?
-Capillary electrophoresis is a method used for separating and identifying fragments of fluorescently labeled DNA by passing them through very thin polymer-filled capillaries.
How does the use of capillaries in capillary electrophoresis differ from conventional slab gels?
-Capillaries allow for the application of a much stronger electric field, resulting in faster separation and higher throughput compared to conventional slab gels.
What are the key features of the capillary electrophoresis process?
-The process features single-based resolution and requires small sample quantities.
What are some applications of capillary electrophoresis in laboratories?
-Applications include Short Tandem Repeat (STR) analysis, Sanger sequencing, and many other everyday applications used by life science and forensic laboratories.
How are fluorescently labeled DNA samples prepared for capillary electrophoresis?
-The samples are prepared and loaded into the capillary electrophoresis instrument, where a capillary array is filled and placed into the sample wells.
What happens when the capillary array enters the sample wells?
-An electric current is formed, and the DNA fragments are moved electrokinetically into the capillaries.
What is the role of the cathode and anode buffer in the process?
-The ends of the capillary array are inserted into the cathode and anode buffer, which helps in controlling the sample migration through the capillary array.
How does the instrument control the sample migration?
-The instrument applies a specific voltage and temperature to control the sample migration through the capillary array.
What is the final step in the capillary electrophoresis process?
-The final step is sample detection, where fluorescently labeled fragments are detected as they move past the detection window.
How are the fluorescent signatures of each fragment recorded?
-As the fragments move past the detection window, the laser excites the fluorescent dyes, and the emitted light is spectrally separated and recorded by a CCD camera.
What happens to the raw data after it is recorded by the CCD camera?
-The raw data is sent to the collection software in the form of a peak and is then ready for analysis using a chosen software program.
Who brought this animation about capillary electrophoresis?
-The animation is brought to you by Spectrum Compact, a company that provides a small but mighty capillary electrophoresis instrument for laboratories.
Outlines
🧬 Capillary Electrophoresis: DNA Fragment Separation
Capillary electrophoresis is a technique used for the separation and identification of fluorescently labeled DNA fragments. This method employs thin polymer-filled capillaries instead of traditional slab gels, allowing for a stronger electric field, which in turn accelerates the separation process and increases the rate of analysis. The process is characterized by single-base resolution and requires minimal sample volumes. It is widely used in STR analysis, Sanger sequencing, and various other applications in life science and forensic laboratories. The script describes the preparation of DNA samples, their loading into the instrument, the application of an electric current to move the DNA fragments into the capillaries, and the subsequent separation based on size. The smallest fragments travel the fastest. The final step involves the detection of the fragments as they pass a detection window, where a laser excites the fluorescent dyes, and the emitted light is recorded by a CCD camera, capturing the fluorescence signature and migration time of each fragment. The data is then sent to collection software for analysis, as demonstrated by the Spectrum Compact CE system.
Mindmap
Keywords
💡Capillary Electrophoresis
💡Fluorescently Labeled DNA
💡Capillaries
💡Electric Field
💡Separation
💡Sample Quantities
💡STR Analysis
💡Sanger Sequencing
💡Laser
💡CCD Camera
💡Collection Software
Highlights
Capillary electrophoresis is a method for separating and identifying fluorescently labeled DNA fragments.
It uses thin polymer-filled capillaries instead of conventional slab gels.
Allows for a stronger electric field, resulting in faster separation and higher throughput.
The process offers single-based resolution and requires small sample quantities.
Applications include STR analysis, Sanger sequencing, and other everyday lab uses.
Fluorescently labeled DNA samples are prepared and loaded into the instrument.
A capillary array is filled and placed into sample wells to form an electric current.
DNA fragments are moved electrokinetically into the capillaries.
The capillary array ends are inserted into cathode and anode buffers.
Instrument applies specific voltage and temperature to control sample migration.
Fragments are separated by size, with the smallest moving the fastest.
Sample detection involves a laser exciting fluorescent dyes and a CCD camera recording the fluorescence.
The CCD camera records the fluorescence signature and migration time of each fragment.
Raw data is sent to collection software as peaks ready for analysis.
The animation is brought to you by Spectrum Compact, a compact CE system for labs.
Spectrum Compact is described as a small but mighty CE instrument.
Transcripts
[Music]
capillary electrophoresis is a method
for separating and identifying fragments
of fluorescently labeled DNA by passing
them through very thin polymer filled
capillaries the use of capillaries
instead of a conventional slab gel makes
it possible to apply a much stronger
electric field resulting in faster
separation and higher throughput the
process features single based resolution
and requires small sample quantities
applications include STR analysis and
Sanger sequencing as well as many other
everyday applications used by life
science and forensic laboratories here's
how it works
first fluorescently labeled DNA samples
are prepared for electrophoresis and
loaded into the seedy instrument a
capillary array is filled a column ER
and placed into the sample wells when
the capillary array enters the wells an
electric current is formed and the DNA
fragments are moved electric kinetically
into the capillaries next the ends of
the capillary array are inserted into
the cathode and anode buffer the
instrument applies a specific voltage
and temperature to control sample
migration through the capillary array
the fragments are separated by size with
the smallest fragments moving and
fastest the final step is sample
detection as a fluorescently labeled
fragments move past the detection window
the laser excites the fluorescent dyes
the emitted light is spectrally
separated and recorded by a CCD camera
the CCD camera records the fluorescence
signature of each fragment as well as
the elapsed time for the fragment to
migrate through the polymer the raw data
is sent to the collection software in
the form of a peak and is now ready for
analysis using your choice of software
program
this animation is brought to you by
spectrum compact seee system a small but
mighty seee instrument for your lab
[Music]
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