From 2013.igem.org
RFC10 Standardization of Tyc Modules
Primer Design
Amplification of Tyc A
TycA | µl
|
Bacillus | 1
|
AT01 | 2
|
AT02 | 2
|
Q5 2x Master mix | 10
|
ddH20 | 5
|
Cycles | temperature [°C] | Time [min:s]
|
1 | 98 | 2:00
|
35 | 98 | 0:10
|
59 | 0:30
|
72 | 1:30
|
1 | 72 | 2:00
|
1 | 10 | inf
|
Result There was a band at the right length but we were not able to cut.
Amplification of Tyc B1
A
TycB1 | µl
|
Bacillus | 1
|
AT03 | 2
|
AT04 | 2
|
Q5 2x Master mix | 10
|
ddH20 | 5
|
Cycles | temperature [°C] | Time [min:s]
|
1 | 98 | 2:00
|
35 | 98 | 0:10
|
56 | 0:30
|
72 | 1:30
|
1 | 72 | 2:00
|
1 | 10 | inf
|
Result no band in gel, empty lane; optimization