Team:TU Delft/2 August 2010 content

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(Difference between revisions)
(→Alkane Sensing, Solvent Tolerance and Salt Tolerance)
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The plates containing yesterday's ligations contained colonies, to check whether they really contain the desired BioBrick a [[Team:TU_Delft/protocols/colony PCR|colony PCR]] was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.
The plates containing yesterday's ligations contained colonies, to check whether they really contain the desired BioBrick a [[Team:TU_Delft/protocols/colony PCR|colony PCR]] was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.
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=Alkane degradation=
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For the next step in BioBrick formation a digestion was done:
 +
Digestion
 +
Some BioBricks are in production. [[Team:TU_Delft/protocols/restriction_enzyme_digestion| Digestion reactions]]:
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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
 +
|'''#'''
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|'''Sample'''
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|'''Enzyme 1'''
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|'''Enzyme 2'''
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|'''Enzyme 3'''
 +
|'''Buffer'''
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|'''BSA'''
 +
|'''Needed fragment'''
 +
|-
 +
|1
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|1 μg 007T 
 +
|EcoRI 
 +
|SpeI
 +
|BamH1
 +
| 2 (Biolabs)
 +
|✓
 +
|‘E – J61100-AlkB2 – S’
 +
|-
 +
|2
 +
|1 μg 008A 
 +
|EcoRI 
 +
|XbaI
 +
|-
 +
|2 (Biolabs)
 +
|✓
 +
|‘E – J61100-rubA3 – X’
 +
|-
 +
|3
 +
|1 μg 010A 
 +
|EcoRI 
 +
|SpeI
 +
|AseI
 +
| 2 (Biolabs)
 +
|✓
 +
|‘E – J61100-rubR – S’
 +
|-
 +
|4
 +
|2 μg B0015 
 +
|EcoRI
 +
|XbaI
 +
|-
 +
| 2 (Biolabs)
 +
|✓
 +
|‘E – B0015 – pSB1AK3 – X’
 +
|-
 +
|5
 +
|1 μg 017A
 +
|EcoRI 
 +
|SpeI
 +
|AseI
 +
|2 (Biolabs)
 +
|✓
 +
|‘E – J61100-ladA – S’
 +
|-
 +
|6
 +
|1 μg 018A
 +
|XbaI 
 +
|PstI
 +
|AseI
 +
|2 (Biolabs)
 +
|✓
 +
|‘X – J61101-ADH – P’
 +
|-
 +
|7
 +
|1 μg 019A
 +
|EcoRI
 +
|SpeI
 +
|AseI
 +
|2 (Biolabs)
 +
|✓
 +
|‘E – J61107-ALDH – S’
 +
|-
 +
|8
 +
|1 μg pSB1K3
 +
|EcoRI
 +
|PstI
 +
|2 (Biolabs)
 +
|✓
 +
|‘E – pSB1K3 – P’
 +
|-
 +
|}
 +
 +
These

Revision as of 09:19, 3 August 2010

Alkane Sensing, Solvent Tolerance and Salt Tolerance

by Pieter

The plates containing yesterday's ligations contained colonies, to check whether they really contain the desired BioBrick a colony PCR was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.

Alkane degradation

For the next step in BioBrick formation a digestion was done: Digestion Some BioBricks are in production. Digestion reactions:

# Sample Enzyme 1 Enzyme 2 Enzyme 3 Buffer BSA Needed fragment
1 1 μg 007T EcoRI SpeI BamH1 2 (Biolabs) ✓ ‘E – J61100-AlkB2 – S’
2 1 μg 008A EcoRI XbaI
2 (Biolabs) ✓ ‘E – J61100-rubA3 – X’
3 1 μg 010A EcoRI SpeI AseI 2 (Biolabs) ✓ ‘E – J61100-rubR – S’
4 2 μg B0015 EcoRI XbaI
2 (Biolabs) ✓ ‘E – B0015 – pSB1AK3 – X’
5 1 μg 017A EcoRI SpeI AseI 2 (Biolabs) ✓ ‘E – J61100-ladA – S’
6 1 μg 018A XbaI PstI AseI 2 (Biolabs) ✓ ‘X – J61101-ADH – P’
7 1 μg 019A EcoRI SpeI AseI 2 (Biolabs) ✓ ‘E – J61107-ALDH – S’
8 1 μg pSB1K3 EcoRI PstI 2 (Biolabs) ✓ ‘E – pSB1K3 – P’

These

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