This protocol explains how a
SpectroCHIP is run on the Mass spec in order to produce
genotypes.
Materials
Spotted SpectroCHIP
Forceps
Method
Insert the chip into the
Mass spectrometer
Push the button on the
computer monitor to toggle to the Sun computer.
Using the right hand mouse,
left-click on the “probe-out button”, and
wait until the light turns green.
Open the probe door and
remove the chip stage out of the Mass spec. Unless
you are placing a chip in immediately, close the probe
door to prevent any dust getting into the chamber.
Any chips found on the
stage should be replaced into their correct packet,
then placed in teh dessicator incase they need to be
rerun
Using the forceps, place
the spotted SpectroCHIPs on the stage, and load the
stage in the Mass spec so that a small amount of the
stage is protruding from the opening, then close the
door to push the remainder of the stage into the machine.
Leave the empty chip packet
next to the Mass spec in the order that you placed the
chips in the machine so that the correct chip can be
replaced into the correctly labeled packet after the
run.
Using the right hand mouse,
left-click on the “probe in button”, and
wait until the light turns green.
Push the button on the
computer monitor to toggle to the RT computer.
Collect data using SpectroTYPER-RT
Login to the RT network
with your own user name and password, if someone else
is logged in then log out and log back in again with
your own log in details.
Make sure that SpectroCaller
is running (look for a Blue "C" in the right hand corner
of the screen ), if not double click on the SpectroCaller
icon.
Using the left mouse,
left-click the SpectroAquire button.
Left-click on the “Auto
Run Set Up” tab.
Enter the chip names in
their corresponding places, the chip names should be
exactly the same as those used to queue the chip in
plate editor, as described in HPGF-SOP014.
Click the high voltage
button on and it will turn red, and check the box that
says turn off HV after last chip is run. Check
that the voltage has reached 19.9 before proceeding.
Click the “Auto
Run tab” and click the “Start Auto Run”
button.
Turn on the monitor that
watches the laser fire on the sample.
Turn the light on behind
the panel next to the probe door.
Watch the calibrant sample
peaks to make sure the machine is working correctly.
There should be three intense peaks and there will be
no sample genotypes produced unless the Mass spec reads
the three calibrant peaks.
Turn SpectroWatcher on
the server computer and add the file name into the file
watched by copying the file name from the notepad icon
called SpectroWatcher and paste it in the file to be
watched column.
Once the file with your
plate name is up on the screen as processed, you can
look at your plate.
Enter your plate in the
Run Log. Start menu, select Programs, Oragen,
Oragen helper, Application, Log in with Generations
user name and password.
Pull down the Tools menu,
select Sequenom Lab Log. Fill out the information.
Safety Implications
All users should receive complete
training in the use of the Brukker Mass Spec before attempting
to use it. Dangers include high voltage and lasers.
No maintenance should be attempted on the Brukker Mass spec
or on software without prior authorization from the Sequenom
Technical Help Advisor.
Related MSDS
None.
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