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Comet Labs WN591 Manual de usuario Pagina 7

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Help! There is a comet in my computer! 4
Figure 1. The principle of fluorescence and absorption and emission spectra of DNA-bound
ethidium bromide. A: DNA-bound ethidium bromide (EB) absorbs green light (high energy) and
emits orange light (low energy). B: DNA-bound ethidium bromide absorbs green light (peak at
~530 nm) and emits orange light (peak at ~610 nm). The excitation filter on the microscope passes
green light and blocks all other wavelengths. The emission filter passes wavelengths longer than
the excitation light wavelength (orange and red). Note that ethidium bromide has another
absorption peak in the UV range of the light spectrum (not shown) – this peak is exploited for
visualisation of DNA on agarose gels with a UV transilluminator. See Figure 2 for the position of
filters in the microscope light path.
0
20
40
60
80
100
400 500 600 700
Wavelength (nm)
Absorption and Emission (a.u.
)
DNA/EtBr absorption
DNA/EtBr emission
excitation filter
emission filter
Wavelength of light (nm)
EB
excitation
(energy absorbed)
p
h
o
t
o
n
400
500
600
700
high energy
EB
emmision
(energy rel eased)
p
h
o
t
o
n
low energy
Energy
high energylow energy
A
B
0
20
40
60
80
100
400 500 600 700
Wavelength (nm)
Absorption and Emission (a.u.
)
DNA/EtBr absorption
DNA/EtBr emission
excitation filter
emission filter
0
20
40
60
80
100
400 500 600 700
Wavelength (nm)
Absorption and Emission (a.u.
)
DNA/EtBr absorption
DNA/EtBr emission
0
20
40
60
80
100
400 500 600 700
Wavelength (nm)
Absorption and Emission (a.u.
)
DNA/EtBr absorption
DNA/EtBr emission
excitation filter
emission filter
Wavelength of light (nm)
EB
excitation
(energy absorbed)
p
h
o
t
o
n
400
500
600
700
high energy
EB
emmision
(energy rel eased)
p
h
o
t
o
n
low energy
Energy
high energylow energy
A
Wavelength of light (nm)
EB
excitation
(energy absorbed)
p
h
o
t
o
n
400
500
600
700
high energy
EB
emmision
(energy rel eased)
p
h
o
t
o
n
low energy
Energy
high energylow energy
Wavelength of light (nm)
EB
excitation
(energy absorbed)
p
h
o
t
o
n
400
500
600
700
high energy
EB
emmision
(energy rel eased)
p
h
o
t
o
n
Wavelength of light (nm)
EBEB
excitation
(energy absorbed)
p
h
o
t
o
n
p
h
o
t
o
n
400
500
600
700
400
500
600
700
high energy
EBEB
emmision
(energy rel eased)
p
h
o
t
o
n
low energy
Energy
high energylow energy
A
B
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