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Team:Groningen/27 June 2013
From 2013.igem.org
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<br/>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 | ||
<br/>0:30 0:10 0:25 0:27 10:00 forever | <br/>0:30 0:10 0:25 0:27 10:00 forever | ||
| + | |||
| + | Added 1 ul serva/100 ml 0.8% agarose gel. | ||
| + | <br/>Gel run at 90V for 22 min at a 0.8% agarose gel. | ||
| + | <br/>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. | ||
Revision as of 11:59, 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
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.
