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
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