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Team:Groningen/26 June 2013
From 2013.igem.org
(Difference between revisions)
| Line 14: | Line 14: | ||
The following protocol is used for the PCR of EstA, MotB and LytB: | The following protocol is used for the PCR of EstA, MotB and LytB: | ||
| - | 98°C, 98°C, 50°C, 72°C, 72°C, 4°C | + | <br/>98°C, 98°C, 50°C, 72°C, 72°C, 4°C |
| - | 0:30 0:10 0:25 0:05 10:00 forever | + | <br/>0:30 0:10 0:25 0:05 10:00 forever |
The following protocol is used for the PCR of FliZ: | The following protocol is used for the PCR of FliZ: | ||
| - | 98°C, 98°C, 45°C, 72°C, 72°C, 4°C | + | <br/>98°C, 98°C, 45°C, 72°C, 72°C, 4°C |
| - | 0:30 0:10 0:25 0:04 10:00 forever | + | <br/>0:30 0:10 0:25 0:04 10:00 forever |
Revision as of 13:33, 26 June 2013
Extraction of the genomic DNA of B. subtilis strain 168.
Diluting the primers (to 100 mM and 10 mM)
Diluting the dNTPs to a concentration of 10 mM.
Run a PCR for the four different signal sequences.
-FliZ
-MotB
-EstA
-LytB
The size will be around 100-150 bp.
The following protocol is used for the PCR of EstA, MotB and LytB:
98°C, 98°C, 50°C, 72°C, 72°C, 4°C
0:30 0:10 0:25 0:05 10:00 forever
The following protocol is used for the PCR of FliZ:
98°C, 98°C, 45°C, 72°C, 72°C, 4°C
0:30 0:10 0:25 0:04 10:00 forever
