Team:Groningen/27 June 2013

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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.