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Team:Groningen/26 June 2013
From 2013.igem.org
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Extraction of the genomic DNA ''B. subtilis''. | Extraction of the genomic DNA ''B. subtilis''. | ||
| - | Diluting the primers. | + | Diluting the primers (to 100 mM and 10 mM) |
| + | <br/>Diluting the dNTPs to a concentration of 10 mM. | ||
| + | |||
| + | Run a PCR for the four different signal sequences. | ||
| + | <br/>-FliZ | ||
| + | <br/>-MotB | ||
| + | <br/>-EstA | ||
| + | <br/>-LytB | ||
| + | <br/>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 | ||
Revision as of 13:27, 26 June 2013
Extraction of the genomic DNA B. subtilis.
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
