4.
Time Mode
— When this mode is selected, the SECONDS LED lights to
indicate that the value displayed or to be entered represents
seconds. Enter the exposure time desired to a maximum of 9,999
seconds (2 hours, 46 minutes and 39 seconds). The minimum value that
can be entered is one second.
The desired time should be entered on the number pad after the ‘‘TIME’’
pad is pressed. Press ‘‘START’’ to begin operation. After the
time has expired, the Spectrolinker beeps four times and the display
flashes ‘‘END.’’ Open the door at this point and the display will oscillate
between ‘‘DOOR’’ and ‘‘0.’’
If you want to repeat the previous operation, just close the door
and press ‘‘START.’’ The Spectrolinker remains in the TIME mode and
displays the last time entered.
To alter a previously entered time, press ‘‘RESET,’’ enter the new value
and then press ‘‘START’’ to begin the operation.
When the chamber door is opened during an operation, the display will
oscillate between ‘‘DOOR’’ and the time remaining. To resume the opera-
tion, close the door and press ‘‘START.’’ Otherwise, press ‘‘RESET’’ to
return to the WAKE state.
APPLICATIONS
CROSSLINKING OF DNA/RNA
The Spectrolinker can be used to covalently bind nucleic acids to mem-
branes after Northern, Southern, slot or dot blotting, and colony or plaque
lifts. Studies [1,2,3] indicate that 120 mJ/cm
2
is an optimal dosage for attach-
ment of DNA or RNA in any of these procedures to nylon or nitrocellulose
membranes. In order to obtain the best results following the DNA or RNA
transfer step, the membrane should be placed inside the Spectrolinker
chamber after the membrane is dried. Place the membrane with the side to
which the nucleic acids are attached facing upwards (i.e. towards the light
source) so that the nucleic acids are directly irradiated. Select an energy
value or press ‘‘OPTIMAL CROSSLINK’’ followed by ‘‘START’’ to initiate the
desired function.
NICKING DNA
The Spectrolinker can be used to nick the ethidium-bromide-stained DNA in
agarose gels in place of a depurination wash [4].
GENE MAPPING
The Spectrolinker can be used to create thymine dimers prior to treatment
with a restriction enzyme. The formation of dimers at or near the recognition
site inhibits cleavage [5].
7
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