Protocol Wizard Biometra TRIO
68 Version 03_19
Figure 47 Biometra TRIO screen for primer concentration adjustment
Press the [EDIT PRIMER] button to toggle to the primer annealing temperature calculator
screen (see Figure 47).
Or press the [CALC. PRIMER] button to calculate the theoretical primer annealing tem-
perature (T
a
).
The calculated values for the forward primer melting temperature (T
m
), reverse primer
melting temperature (T
m
), mean primer melting temperature (T
m
) and calculated
annealing temperature (T
a
) are displayed.
Press the [SAVE TEMP.] button to transfer the calculated primer annealing temperature
to the protocol wizard (see Figure 46).
The calculated T
a
value will be displayed in the input box for the primer annealing
temperature.
Press the [INFO] button to get information on the algorithms used for annealing tem-
perature calculations.
For short sequences of 14 nucleotides or less the software calculates the primer an-
nealing temperature (T
a
) based on the primer T
m
values as follows:
1. For each primer the melting temperature is calculated using the following equation
according to Wallace et al.
1
:
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