Team:Groningen/Labwork/16 July 2013

From 2013.igem.org

(Difference between revisions)
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'''Inne'''
'''Inne'''
<br> Did a PCR with 3 samples in duplo
<br> Did a PCR with 3 samples in duplo
-
<br>sample 1: primer F without strep tag
+
<table border="1">
-
<br>primer R without strep tag
+
<tr>
-
<br>annealing temp: 78 C
+
<td>Sample</td>
-
<br>sample 2: primer F with strep tag
+
<td>Primer F</td>
-
<br>primer R without strep tag
+
<td>Primer R</td>
-
<br>annealing temp 78 C
+
<td>Annealing temp.</td>
-
<br>sample 3: primer F without strep tag
+
</tr>
-
<br>primer R with strep tag
+
<tr>
-
<br>annealing temp 80 C
+
<td>1</td>
 +
<td>without strep tag</td>
 +
<td>without strep tag</td>
 +
<td>78 C</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
 +
<td>with strep tag</td>
 +
<td>without strep tag</td>
 +
<td>78 C</td>
 +
</tr>
 +
<tr>
 +
<td>3</td>
 +
<td>without strep tag</td>
 +
<td>with strep tag</td>
 +
<td>80 C</td>
 +
</tr>
 +
</table>
Protocol:
Protocol:
-
<br>28.2 uL MillQ
+
<table border="1">
-
1.5 uL 5% DMSO
+
<tr>
-
1 uL of each dNTP
+
<td>amount</td>
-
10 uL GC buffer
+
<td>compound</td>
-
2.5 uL primer F
+
</tr>
-
2.5 uL primer R
+
<tr>
-
1 uL template DNA bba_16 (Utah silk biobrick)
+
<td>28.2 uL</td>
-
0.3 uL phusion polymerase
+
<td>MilliQ</td>
 +
</tr>
 +
<tr>
 +
<td>1.5 uL</td>
 +
<td>5% DMSO</td>
 +
</tr>
 +
<tr>
 +
<td>1 uL</td>
 +
<td>each dNTP</td>
 +
</tr>
 +
<tr>
 +
<td>10 uL</td>
 +
<td>Phusion GC buffer</td>
 +
</tr>
 +
<tr>
 +
<td>2.5 uL</td>
 +
<td>Primer F</td>
 +
</tr>
 +
<tr>
 +
<td>2.5 uL</td>
 +
<td>Primer R</td>
 +
</tr>
 +
<tr>
 +
<td>1 uL</td>
 +
<td>Silk template BBa_16</td>
 +
</tr>
 +
<tr>
 +
<td>0.3 uL</td>
 +
<td>Phusion polymerase</td>
 +
</tr>
 +
</table>

Revision as of 09:58, 16 July 2013

Mirjam
Finally the colonies are growing on LB agar plates with ampicillin. The negative control does not show any colonies, the double negative does??, the positive control shows a lot of red colonies. On one of the four plates there are no colonies present, on two of them only a couple and on one a lot.

The colonies are picked and placed in LB medium with ampicillin to grow further for miniprep.

Inne
Did a PCR with 3 samples in duplo

Sample Primer F Primer R Annealing temp.
1 without strep tag without strep tag 78 C
2 with strep tag without strep tag 78 C
3 without strep tag with strep tag 80 C

Protocol:

amount compound
28.2 uL MilliQ
1.5 uL 5% DMSO
1 uL each dNTP
10 uL Phusion GC buffer
2.5 uL Primer F
2.5 uL Primer R
1 uL Silk template BBa_16
0.3 uL Phusion polymerase


Sander
did a Restriction digestion for Silk and Signal Sequences. incubation started at 11:00 for the signal sequences and 11:30 for the silk.

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