Team:Heidelberg/Tyrocidine week17 biobrickmod

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Revision as of 21:34, 2 October 2013

Contents

RFC10 Standardization of Tyc Modules

Tyc C5

what µl
Bacillus 1
AT05 2
AT06 2
Q5 2x Master mix 10
ddH20 5
Cycles temperature [°C] Time [s]
1 98 2:00
35 98 10
61 0:30
72 1:00
1 72 2:00
1 10 inf

Tyc C6

what µl
Bacillus 1
AT07 2
AT08 2
Q5 2x Master mix 10
ddH20 5
Cycles temperature [°C] Time [s]
1 98 2:00
35 98 10
56 0:30
72 1:00
1 72 2:00
1 10 inf

Tyc A 2013-08-20

Since neither Tyc B1, C5, C6 where succesfully amplified and Tyc A was rather a light band we decided to first find the optimal protocol for Tyc A. Also the Q5 Polymerase was contaminated we decided to use Phusion Flash which is faster and cheaper than the Q5.

Optimiziation A

Reaction
what µl
Bacillus 1
AT01 2
AT02 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 5
Condition I
Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:01
56 0:05
72 0:55
1 72 5:00
1 10 inf
Conditions II
Cycles temperature [°C] Time [min:s]
1 98 2:00
14 98 0:01
59 ↓0.5 0:05
72 0:55
16 98 0:01
56 0:05
72 0:55
1 72 5:00
1 10 inf
Conditions III
Cycles temperature [°C] Time [min:s]
1 98 2:00
30 98 0:01
72 0:55
1 72 5:00
1 10 inf

Tyc B1 2013-08-21

Reaction
what µl
Bacillus 1
AT03 2
AT04 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 5
Condition
Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:01
56 0:05
72 0:55
1 72 5:00
1 10 inf

Tyc C5 2013-08-21

what µl
Bacillus 1
AT05 2
AT06 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 5
Condition
Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:01
61 0:05
72 0:55
1 72 5:00
1 10 inf

Tyc C6 2013-08-21

what µl
Bacillus 1
AT07 2
AT08 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 5
Condition
Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:01
56 0:05
72 0:55
1 72 5:00
1 10 inf

Tyc B1 2013-08-22

Reaction
what µl
Bacillus 1
AT03 2
AT04 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 5


Conditions II
Cycles temperature [°C] Time [min:s]
1 98 2:00
14 98 0:01
63 ↓0.5 0:05
72 0:55
16 98 0:01
58 0:05
72 0:55
1 72 5:00
1 10 inf

Tyc C5 2013-08-22

Reaction
what µl
Bacillus 1
AT05 2
AT06 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 4


Conditions II
Cycles temperature [°C] Time [min:s]
1 98 2:00
14 98 0:01
61 ↓0.5 0:05
72 0:55
16 98 0:01
63 0:05
72 0:55
1 72 5:00
1 10 inf

Tyc B1 2013-08-23

B 23-08-2013

Reaction
what µl
Bacillus 1
AT03 2
AT04 2
Phusion Flash 2x Master Mix 10
DMSO 1.5
ddH20 3.5


Conditions
Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:01
63 0:05
72 0:50
1 72 5:00
1 10 inf

Tyc C5 2013-08-23

B 23-08-2013

Reaction I
what µl
Bacillus 1
AT05 2
AT06 2
Phusion Flash 2x Master Mix 10
DMSO 1.5
ddH20 3.5
Reaction II
what µl
Bacillus 1
AT05 2
AT06 2
Phusion Flash 2x Master Mix 10
DMSO 1
ddH20 4


Conditions
Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:01
63 0:05
72 0:50
1 72 5:00
1 10 inf
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