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).
E
DIT PRIMER
Or press the [
CALC
.
PRIMER
] button to calculate the theoretical primer annealing tem-
perature (T
a
).
C
ALC
.
PRIMER
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).
S
AVE TEMP
.
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.
I
NFO
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
: