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Team:Groningen/27 June 2013
From 2013.igem.org
(Difference between revisions)
| Line 1: | Line 1: | ||
Today the PCR products of the signal sequences (FliZ, EstA, MotB and LytB) are purified using gel purification. | Today the PCR products of the signal sequences (FliZ, EstA, MotB and LytB) are purified using gel purification. | ||
| - | A PCR is done for some different silk constructs to start with. The following combinations are made: | + | A PCR is done for some different silk constructs to start with. The following combinations are made (same protocol as 26-06-2013): |
<br/>1) signal sequence - N-terminal strep tag - silk | <br/>1) signal sequence - N-terminal strep tag - silk | ||
<br/>2) signal sequence - silk - C-terminal strep tag | <br/>2) signal sequence - silk - C-terminal strep tag | ||
| Line 8: | Line 8: | ||
The following protocol is used: | The following protocol is used: | ||
| + | <br/>98°C, 98°C, 50°C, 72°C, 72°C, 4°C | ||
| + | <br/>0:30 0:10 0:25 0:27 10:00 forever | ||
Revision as of 11:04, 27 June 2013
Today the PCR products of the signal sequences (FliZ, EstA, MotB and LytB) are purified using gel purification.
A PCR is done for some different silk constructs to start with. The following combinations are made (same protocol as 26-06-2013):
1) signal sequence - N-terminal strep tag - silk
2) signal sequence - silk - C-terminal strep tag
3) signal sequence - silk - no strep tag
4) silk without tag
The following protocol is used:
98°C, 98°C, 50°C, 72°C, 72°C, 4°C
0:30 0:10 0:25 0:27 10:00 forever
