Team:TU Delft/4 August 2010 content

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=Alkane Sensing, Solvent Tolerance and Salt Tolerance=
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=Alkane Sensing, Solvent Tolerance, Salt Tolerance, Emulsifier=
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==Ligation==
 
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==Transformation==
 
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=Emulsifier=
 
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While waiting for the sequencing results of BBa_K398202 we already proceed with the insertion of OprG and AlnA.
 
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Digestion
 
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Ligation
 
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Transformation
 

Revision as of 14:44, 5 August 2010

Alkane Sensing, Solvent Tolerance, Salt Tolerance, Emulsifier

by Pieter

We found it remarkable that the only positive samples we found so far were from the series without preamplifying the insert by PCR (e.g. the AlkS). This is why we decided to retry the insertion of bbc1, PhPFDalpha and PhPFDbeta in J61101, but this time without PCRing the insert.

Digestion

For the assembly of BBa_K398101, BBa_K398205, BBa_K398206, BBa_K398402 and BBa_K398403 a digestion was done:

# Sample Enzyme 1 Enzyme 2 Enzyme 3 Buffer BSA Needed fragment
1 1 μg bbc1 XbaI PstI ScaI 2 (Biolabs) ✓ ‘X – bbc1 - P’
2 1 μg PhPFDalpha XbaI PstI ScaI 2 (Biolabs) ✓ ‘X – PhPFDalpha - P’
3 1 μg PhPFDbeta XbaI PstI ScaI 2 (Biolabs) ✓ ‘X – PhPFDbeta - P’
4 1 μg J61101 SpeI PstI 2 (Biolabs) ✓ ‘S – J61101 - P’
5 1 μg OprG-B0015 XbaI PstI 2 (Biolabs) ✓ ‘X – OprG-B0015 - P’
6 1 μg AlnA-B0015 XbaI PstI 2 (Biolabs) ✓ ‘X – AlnA-B0015 - P’
7 1 μg R0011-B0032 (sample 9) SpeI PstI 2 (Biolabs) ✓ ‘S – R0011-B0032 - P’
8 1 μg R0011-B0032 (sample 10) SpeI PstI 2 (Biolabs) ✓ ‘S – R0011-B0032 - P’
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