Team:Groningen/Labwork/16 July 2013

From 2013.igem.org

(Difference between revisions)
Line 5: Line 5:
'''Inne'''
'''Inne'''
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<br> Did a PCR with 3 samples
+
<br> Did a PCR with 3 samples in duplo
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sample 1: primer F without strep tag
+
<br>sample 1: primer F without strep tag
-
primer R without strep tag
+
<br>primer R without strep tag
-
annealing temp: 78 C
+
<br>annealing temp: 78 C
-
sample 2: primer F with strep tag
+
<br>sample 2: primer F with strep tag
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primer R without strep tag
+
<br>primer R without strep tag
-
annealing temp 78 C
+
<br>annealing temp 78 C
-
sample 3: primer F without strep tag
+
<br>sample 3: primer F without strep tag
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primer R with strep tag
+
<br>primer R with strep tag
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annealing temp 80 C
+
<br>annealing temp 80 C
 +
Protocol:
 +
<br>28.2 uL MillQ
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1.5 uL 5% DMSO
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1 uL of each dNTP
 +
10 uL GC buffer
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2.5 uL primer F
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2.5 uL primer R
 +
1 uL template DNA bba_16 (Utah silk biobrick)
 +
0.3 uL phusion polymerase

Revision as of 09:38, 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 1: primer F without strep tag
primer R without strep tag
annealing temp: 78 C
sample 2: primer F with strep tag
primer R without strep tag
annealing temp 78 C
sample 3: primer F without strep tag
primer R with strep tag
annealing temp 80 C

Protocol:
28.2 uL MillQ 1.5 uL 5% DMSO 1 uL of each dNTP 10 uL GC buffer 2.5 uL primer F 2.5 uL primer R 1 uL template DNA bba_16 (Utah silk biobrick) 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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