"
Team:Groningen/26 June 2013
From 2013.igem.org
(Difference between revisions)
| Line 3: | Line 3: | ||
Diluting the primers (to 100 mM and 10 mM) | Diluting the primers (to 100 mM and 10 mM) | ||
<br/>Diluting the dNTPs to a concentration of 10 mM. | <br/>Diluting the dNTPs to a concentration of 10 mM. | ||
| + | |||
Run a PCR for the four different signal sequences. | Run a PCR for the four different signal sequences. | ||
| Line 10: | Line 11: | ||
<br/>-LytB | <br/>-LytB | ||
<br/>The size will be around 100-150 bp. | <br/>The size will be around 100-150 bp. | ||
| + | |||
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: | ||
Revision as of 13:28, 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
