user manual

blotting equipment

Hoefer UVC 500

Ultraviolet Crosslinker
transilluminator

um UVC500-IM/Rev. F0/04-04
Page finder

UV Crosslinker function and description . . . . . . . . . . . . . .1

Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2

Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3

Important safety information . . . . . . . . . . . . . . . . . . . . . . . . . .4

Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5

Energy setting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6

Time setting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8

Care and maintenance ............................. 10

Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

Ordering information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

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 UV Crosslinker function and description
The Hoefer™ UVC 500 Ultraviolet Crosslinker
provides a uniform and intense source of ultra-
violet (UV) light for such uses as:
• DNA crosslinking
• DNA nicking
• Rapid restriction site mapping
• Testing RecA function
• Ultraviolet sterilization
• Elimination of PCR contamination

The internal photo-feedback system continually

measures the energy delivered within the chamber
and automatically deactivates the UV light source
once the programmed energy level is reached.
This system also automatically adjusts to varia-
tions in UV intensity that occur as lamps age.

Fig 1. Crosslinker front view.

Pull the door open along this handle LED display

View window Keypad Power switch

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 Unpacking
Unwrap all packages carefully and compare
contents with the packing list, making sure all
items arrived. If any part is missing, contact
your local sales office. Inspect all components
for damage that may have occurred while the
unit was in transit. If any part appears damaged,
contact the carrier immediately. Be sure to keep
all packing material for damage claims or to use
should it become necessary to return the unit.

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Specifications

Ultraviolet wavelength and sources

• 254 nm (shortwave)
• Five 8-W 254 nm UV dual bi-pin discharge
type tubes

Features
• Microprocessor controlled UV sensor feedback
system
• Large LED readout
• Tactile membrane switch keypad
• Laydown-type door
• Multiple set functions: pre-set UV energy or time
exposure user-specified UV
energy or time exposure
• Max. energy setting 999,900 µJ/cm2
• Max. time setting: 999.9 min

Dimensions (h × d × w)
External 22.2 × 34.9 × 40.0 cm
(8.75 × 13.75 × 15.75 in.)
Chamber interior 12.7 × 30.5 × 25.4 cm
(5 × 12 × 10 in.)

Input fuses
• 115 V - F 1A/250V 5 × 20 mm (2 fuses)
• 230 V - F 1A/250V 5 × 20 mm (2 fuses)

Weight
• 7.5 kg (15.5 lbs)

Safety
• Safety interlocked door
• UV-blocking view window
• Safety-fused and removable power cord

Safety certifications
CE 89/336/EEC (EMC directive), CE 73/23/EEC (LV
directive), EN-61010-1 (IEC1010-1, UL3101-1,
CSA22.2 1010-1)

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 Important safety information
• Always wear protective gloves and goggles when
working with the crosslinker.

• The UVC 500 is a powerful source of ultravio-

let radiation. The door is safety interlocked to
prevent accidental user exposure to UV light.
Do not attempt to override the safety interlocks.

If the safety interlock is disengaged, the UV

lamps do not light and the unit will beep 3 times
when the START key is pressed.

If a malfunction occurs and the lamps in the

crosslinker remain on when the door is open,
immediately close the door and turn off the
mains power.

• The view window blocks UV radiation and

allows viewing while the unit is in operation.
If the window is broken or removed, stop using
the crosslinker immediately.

• Always disconnect the mains power cord before

cleaning the unit or replacing fuses or lamps.

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 Operating instructions
Set up and membrane placement

1
Place the unit on a stable and level surface.

2
Note: To protect the cross- Plug the power cord into the receptacle at the back of
linker from corrosive buffers, the unit and into a properly grounded electrical outlet
lay a piece of plastic wrap of the proper power rating.
on the compartment bottom
before placing the blotting 3
paper and sample. Flip the power mains switch located under the
keypad to on (–).
Note: The instrument settings are always the values
last programmed, and are indicated by the LED
display and a lit red indicator lamp.

4
Open the door by pulling the door handle above the
window toward you.
Note: Do not obstruct or damage the photofeedback
sensor, which is mounted on a small metal tab
protruding from the lower right side.

5
Moisten (do not wet completely) a sheet of blotting
paper with transfer buffer and place it in the center
of the compartment. Place the membrane on the
paper with the nucleic acid side facing upward.

6
Set the energy exposure (energy or time) as described
in the next section.

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 Energy setting
The amount of energy recommended for UV
crosslinking of DNA and RNA by covalently
binding nucleic acids to Hybond™ membranes
after Northern, Southern, slot or dot blotting
is 70,000 µJ/cm2. This value is preset at the
factory and displays as 700 on the LED display,
Note: Multiply all displayed
which only shows 4 digits. As the unit operates,
energy settings by 100 for a countdown of the remaining energy to be emit-
the actual energy value. ted is displayed on the LED.

UVC 500 Crosslinker Using the preset energy setting

user interface
1
The LED display shows values for First press the PRESET key and then the ENERGY
energy (multiply by 100 for actual
key. The red indicator lamp next to each of these
value in µJ/cm2) and time (in
minutes and tenths of a minute.) keys should light and the LED should display 700
(or the value last programmed).

2
Press the green START key. The UV lamps take a few
seconds to energize before the countdown value is
displayed. At the end of the exposure, the unit auto-
matically stops and beeps 5 times.

Reprogramming the preset energy setting

1
Press and hold the PRESET key until you hear a
second beep. The lamp next to the PRESET key
flashes when this key switches to program mode.

2
Press the ENERGY key. Use the numeric keypad to
enter the new value. The LED display should show
the new value.

3
Press the ENTER key.

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Programming the ENERGY key

1
Press the ENERGY key.

2
Enter a value using the numeric keypad. The LED will
display the new value.

3
Press ENTER. After programming, pressing the
ENERGY key retrieves this setting. Change the setting
by entering a new value and pressing the ENTER key.

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 Time setting
The display shows time in minutes and tenths of
a minute.

Using the preset time setting

1
First press the PRESET key and then the TIME key.
The red indicator lamp next to each of these keys
should light and the LED should display 5.0 (or the
value last programmed).

2
Press the green START key. The UV lamps take a few
seconds to energize before the countdown value is
displayed. At the end of the exposure, the unit auto-
matically stops and beeps 5 times.

Reprogramming the preset time setting

1
Press and hold the PRESET key until you hear the
second beep. The lamp next to the PRESET key
flashes when this key switches to program mode.

2
Press the TIME key. Use the numeric keypad to enter
the new value. The LED display shows the new value.

3
Press the ENTER key.

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Programming the TIME key

1
Press the TIME key.

2
Enter a value using the numeric keypad. The LED will
display the new value.

3
Press the ENTER key. After programming, press-
ing the TIME key retrieves this setting. Change the
setting by entering a new value and pressing ENTER.

Stopping and restarting the crosslinker

• Press the red STOP/RESET key to stop the
exposure. The LED will display the remaining
exposure level or time.

• Press the green START key to restart the expo-

sure. The program will continue from where it
was stopped.

• To enter a different value once the program has

started, press STOP/RESET, enter the new value,
then press ENTER. Press the START key to begin
the new exposure value.

• To revert to the last used setting after the expo-

sure is interrupted, press the STOP/RESET key a
second time. Press START to start exposure from
the previous setting.

• If the door is accidentally opened during expo-

sure, the UV lamps instantly turn off and the
remaining exposure level or time is displayed on
the LED display. To restart, close the door and
press the START key.

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 Care and maintenance
Cleaning
Preventative maintenance includes regular clean-
Caution! Always turn the ing of the compartment surface, the sensor, and
mains power switch off and UV lamps.
unplug the instrument
before cleaning. • Use mild soap and water on a soft cloth or
sponge to clean the exterior.
• Gently wipe the photofeedback sensor and UV
lamps regularly with a soft cotton cloth damp-
ened with alcohol.

• Always wipe any water and spills from the inside

of the unit and clean away all chemicals after
each use.

• Never use abrasive cleaners, acetone or chloro-

form on any part.

Important! The photofeed- Replacing UV lamps

back system is calibrated
Periodically check that all five lamps function
to 254 nm UV light. Do not
install lamps rated for a by looking through the view window while the
different wavelength. unit is on. If one or more lamps are burned out,
it is recommended that all five be replaced at the
same time. To replace each lamp:

1
Turn off the mains power switch and disconnect the
power cord.

2
Locate the lamps along the top of the compartment.
Remove each lamp by carefully rotating it 1/4 – 1/3
turn and then carefully pulling it loose.

3
To insert the new lamp, hold it so that the metal
plugs at the ends align vertically. Place the lamp in
the sockets at each end of the unit and lightly push it
up into place. Rotate the lamp to lock it into place.

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 Replacing fuses
Fuses protect equipment by disconnecting loads
too large for the instrument’s circuit design. For
continued protection, only use fuses with the
specified voltage and current rating.

Each of two round modules on the left side of

the control panel (above the mains power cord
receptacle) holds one F 1A/250V 5 × 20 mm
(Fast blo) fuse.

1
Caution! Set the mains power switch to off and
detach the power cord before replacing fuses.

2
Insert a small flat-blade screwdriver into the slot on
the fuse module and turn it 1/4 turn counterclock-
wise. The spring-loaded module cap will loosen and
you can then pull the cap/fuse holder out.

3
Pull the fuse out of its holder and inspect. If the fuse
element is burned or broken, replace the fuse with an
identical type. If the fuse appears to be intact, check
it with an ohmmeter.

4
Insert a good fuse into the holder and then insert
this assembly back into the unit. Seat the module by
inserting the screwdriver into the slot, pressing gently,
Fig 2. UVC 500 Crosslinker back and turning the cap 1/4 turn clockwise.
view 115 V model.

Model ID and power specifications

Fuse modules (2)

Mains power cord receptacle

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 Bibliography

DNA crosslinking
Church, M. C. and G. Gilbert. (1984) Genomic
sequencing. Proc Natl. Acad. Sci., USA. 81,
1991–1995.

Khandjian, E. W. (1987) Optimized hybridiza-

tion of DNA blotted and fixed to nitrocellu-
lose and nylon membranes. BioTechnology. 5,
165–167.

Khandjian, E. W. (1986) UV crosslinking of

RNA to nylon membrane enhances hybridiza-
tion signals. Molec. Biol. Report. 11, 107–115.

Reed, K. C. and D. A. Mann. (1985) Rapid

transfer of DNA from agarose gels to nylon
membranes. Nuc. Acid Res. 13, 7207–7221.

DNA nicking
Vollrath, D. and R. W. Davis. (1987) Resolution
of DNA molecules greater than 5 megabases by
contour-clamped homogeneous electric fields.
Nuc. Acid Res. 15, 7865–7876.

Rapid restriction site mapping

Whittaker, P. A. and E. M. Southern. (1986)
Ultraviolet irradiation of DNA: A way of gener-
ating partial digests for rapid restriction site
mapping. Gene. 41, 129–134.

Testing RecA function

Maniatis, T. (1989) Maintenance and Testing of
Lysogens of Bacteriophage Lambda in Molecular
Cloning: A Laboratory Manual, eds. Sambrook,
Fritsch and Maniatis. 2.96.

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Ultraviolet sterilization
Reich, R. R., and Anderson, H. D. (1986) Ster-
ilization of Membrane Filters with Ultraviolet
Radiation. Pharmaceutical Manufacturing. Jan.

Elimination of PCR contamination

WIlson, K. H. (1992) UV Absorption Compli-
cates PCR Decontamination, BioTechniques. 13,
208–210.

Sarkar, Gobinda, and Sommer, Steve S. (1991)

Parameters Affecting Susceptibility of PCR
Contamination to UV Inactivation, BioTech-
niques, 10, 590–594.

Sarkar, Gobinda, and Sommer, Steve S. (1990)

Shedding Light on PCR Contamination. Nature.
343, 27.

Yamada, H. and Hieda, K. (1992) Wavelength

Dependence (150-290nm) of the Formation of
the Cyclobutane Dimer and the (6-4) Photoprod-
uct of Thymine. Photochemistry and Photobiol-
ogy. 55, 541–548.

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 Ordering information
code
product qty. number

Hoefer UVC Ultraviolet Crosslinker

For membranes to 25 × 30 cm.
115 VAC 1 UVC500-115V
230 VAC 1 UVC500-230V
Replacement UV lamp, 8 W, 254 nm 1 UVLS-8
Replacement fuses 5 PSF1A-F8-5X20

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Hoefer, Inc.
953 Indiana Street
San Francisco, CA
94107 USA

www.hoeferinc.com

© 2004 Hoefer, Inc.—

All rights reserved.
Printed in the USA

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