68
•
Temperature:
Set using the sliding thermometer or type a value in the box.
•
Hold Time:
Shown in hrs:min:sec up to a maximum of 99:59:59 (minimum 00:00:01). The
default setting is
seconds
, therefore for a time in seconds, type in the number of seconds
and press the enter key. To set the time in hours, add “h” after the numerical value, e.g. 1h,
and for minutes add “m”.
•
Number of Reads:
Displays the number of readings programmed for the step (maximum of
4).
•
Add Read:
Fluorescence reads can be added by clicking on the
Add Read
button. See
section 3.4.3.5 for details of adding a read to a step.
•
Inc/Dec Temp:
Used in programs where an incremental increase or decrease in
temperature is required. This function is useful in ‘Touchdown’ PCR, for example, where the
annealing temperature is kept high in the initial cycles and then gradually brought down with
the aim of reducing non-specific amplification.
•
Inc/Dec Time:
Used in programs where an incremental increase or decrease in time with
each cycle is required such as long template PCR.
•
Max Heating Rate
: Allows the heating rate to be set between 0.1°C/sec and 2.2°C/sec.
Note: If any special parameters are selected, including Inc/Dec Temp or Inc/Dec Time, then a blue
triangle will appear in the temperature profile plot.
To delete a step, right click on the step and select
Delete
. Alternatively, highlight the step and
select
Delete
from the
Tools
menu in the tool bar.
3.4.3.5
Adding a read to a step
The user must tell the instrument at which point in the thermal cycling program to collect
fluorescent readings and which filter cartridges to use. Reads can either be added at the same
time as the step parameters or added stage-by-stage after the thermal cycle program has been
completed.
•
To add a read to a step, highlight the step to display the step parameters box and click on
Add Read
.
The filter programming options will then be displayed:
•
Select Cartridge by Name/Wavelength
: This is the name given to the cartridge on filter
installation and is stored in the instrument memory. The filter cartridge can either be chosen
on the basis of its name (as given during filter installation) or by its ID wavelength. Click to
