Team:Groningen/27 June 2013

From 2013.igem.org

(Difference between revisions)
Line 10: Line 10:
<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.