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Team:Groningen/27 June 2013
From 2013.igem.org
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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. | ||
| - | FliZ 9,2 ng/ul | + | <br/>FliZ 9,2 ng/ul |
| - | LytB 8,2 ng/ul | + | <br/>LytB 8,2 ng/ul |
| - | MotB 8,8 ng/ul | + | <br/>MotB 8,8 ng/ul |
| - | EstA 8,8 ng/ul | + | <br/>EstA 8,8 ng/ul |
A PCR is done for some different silk constructs to start with. The following combinations are made (same protocol as 26-06-2013): | A PCR is done for some different silk constructs to start with. The following combinations are made (same protocol as 26-06-2013): | ||
Revision as of 12:20, 27 June 2013
Today the PCR products of the signal sequences (FliZ, EstA, MotB and LytB) are purified using gel purification.
FliZ 9,2 ng/ul
LytB 8,2 ng/ul
MotB 8,8 ng/ul
EstA 8,8 ng/ul
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
Added 1 ul serva/100 ml 0.8% agarose gel.
Gel run at 90V for 22 min at a 0.8% agarose gel.
Gel imaging revealed that no bands where present. Therefore it is placed at a high concentration of Ethidium Bromide. The bands appeared. A big smear is seen for all the silk products with a bigger band around 200 bp.
