This document discusses gel documentation, which is a system used to image and document nucleic acid and protein gels stained with fluorescent dyes. It consists of a UV light source, hood, and camera. Fluorescent dyes like ethidium bromide bind to nucleic acids and emit light under UV exposure, with brighter bands indicating more material. The system photographs gels, prints images, and saves files for future use. Components include the camera, lenses, filters, illuminator, stage, and darkroom. Quantity One software is used to operate the system, capturing and analyzing images through tools for detection, measurement, and report generation. Gel documentation has applications in electrophoresis, blotting, and other molecular biology techniques.
2. INTRODUCTION
Gel documentation , also known as gel doc. system , gel image
system , or gel imager.
It is widely used in molecular biology laboratories for imaging and
documentation of nucleic acid and protein polyacrylamide or
agarose gels typically stained with ethidium bromide or flurophores
such as SYBR green.
Generally , a gel doc. is composed of ultraviolet ( UV ) light
transilluminator , a hood to shield external light sources and CCTV
camera for image capturing.
3. PRINCIPLE
Principle of fluorescence with fluorescent staining of
nucleic acids , a fluorescent substance that has bound to
nucleic acid is exited by ultraviolet irradiation and emits
fluorescent light.
Ethidium bromide binds specifically to nucleic acid and
the amount of bonding depends on the molecular weight
and concentration of the nucleic acid.
In other words , a band for a large amount will shine
brighter ; conversely , fluorescence will be weaker for a
band for a small amount.
4. PURPOSE AND ANALYSIS
Photography of stained gel.
Print out of photographic data.
Image data is displayed in
real time.
Image still displayed can be printed
out with a video printer or saved to a
compact flash media for future use.
5. COMPONENTS OF GEL
DOC. SYSTEM
Camera
Lenses
Filters
Overhead illuminator
Visible light
Adjustable stage
Darkroom
Tr ansilluminator
6. COMPONENTS OF GEL
DOC. SYSTEM
Camera : Ultraviolet camera ranging from 1.4m upto
8.3m
pixel.
Lenses : All lanses are computer controlled and motor
driven. Here lances automatically tracks upwards or
downwards movements of sample using auto focus.
Filters : There is an extensive range of emission filters
use for an array of applications.
Over head illuminator : White light , Ultra violet light ,
and LED lighting option
Visible light : For extending the transmitted light
application. Visible light convert screen for visible light
application.
7. COMPONENTS OF GEL
DOC. SYSTEM
Adjustable stage : This enables samples to be moved
closer to the camera.
Darkroom
: dark room is suitable for advanced
chemiluminescence, fluorescence and visible light
application. An electronic auto-door lock
with security
function prevents interruption to long exposures.
Transilluminator : It has a variable intensity setting and
has a safety cut-off.
8. QUANTITY ONE FROM
BIORED
It is a software which is required to operate gel doc. system.
It is user friendly version windows based option.
It has a various tool for image capturing and transformation.
There is a separate tool for identifying , detecting and drawing lense and bands.
Band attributes include band creation , GAUSS model bands , density of a
band , band boundries etc……
There is a separate function for storing standard data , standard curve for
known molecular weight and size.
A separate volume analysis tool for volume contour , free hand , rectangle ,
circle and array.
The software also has options for VNTR display , VNTR
calculation ,
differential display and colony counting applictions.
The software can generate reports for individual bands , all lenses , band types
, 1-dimension analysis , lane compaing , phylogenetic tree and other simmilarity
matrix options.