Team:TU Delft/10 August 2010 content

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=Lab work=
=Lab work=
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==Tolerance and Sensing==
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A new attempt to construct parts 304 (PalkS12 + B0032), 327(P(caif) + B0032) and 405 (402 + 403).
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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
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|'''#'''
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|'''Sample'''
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|'''Enzyme 1'''
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|'''Enzyme 2'''
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|'''Enzyme 3'''
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|'''Buffer'''
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|'''BSA'''
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|'''Needed fragment'''
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|-
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|1
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|PalkS12A 
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|EcoRI 
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|SpeI
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|ScaI
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|2 (Biolabs)
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|✓
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|‘E - PalkS12 - S’
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|-
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|2
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|P(caif)A 
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|EcoRI 
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|SpeI
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|ScaI
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|2 (Biolabs)
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|✓
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|‘E - P(caif) - S’
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|-
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|3
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|B0032
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|EcoRI 
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|XbaI
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|
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|2 (Biolabs)
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|✓
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|‘X - B0032 - E’
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|-
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|4
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|402: J61101 - PhPFDalpha 
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|EcoRI 
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|SpeI
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|ScaI
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|2 (Biolabs)
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|✓
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|‘E - J61101 - PhPFDalpha - S’
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|-
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|5
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|403: J61101 - PhPFDbeta 
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|EcoRI 
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|XbaI
 +
|
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|2 (Biolabs)
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|✓
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|‘E - J61101 - PhPFDbeta - S’
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|}
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==Alkane degradation==
==Alkane degradation==

Revision as of 12:16, 16 August 2010

Lab work

Tolerance and Sensing

A new attempt to construct parts 304 (PalkS12 + B0032), 327(P(caif) + B0032) and 405 (402 + 403).

# Sample Enzyme 1 Enzyme 2 Enzyme 3 Buffer BSA Needed fragment
1 PalkS12A EcoRI SpeI ScaI 2 (Biolabs) ✓ ‘E - PalkS12 - S’
2 P(caif)A EcoRI SpeI ScaI 2 (Biolabs) ✓ ‘E - P(caif) - S’
3 B0032 EcoRI XbaI 2 (Biolabs) ✓ ‘X - B0032 - E’
4 402: J61101 - PhPFDalpha EcoRI SpeI ScaI 2 (Biolabs) ✓ ‘E - J61101 - PhPFDalpha - S’
5 403: J61101 - PhPFDbeta EcoRI XbaI 2 (Biolabs) ✓ ‘E - J61101 - PhPFDbeta - S’

Alkane degradation

All of yesterday's plates contained colonies. To check whether they contain the correct insert we will do a colony PCR. TUDelft 20100810 PCR.png

Lane description

# Description Primers Expected length (bp) ✓ ✗
L SmartLadder n/a n/a n/a n/a
1-5 013A G00100 + G00101 1929 none all
6-10 013K G00100 + G00101 1929 9 6,7,8,10
11-15 020A G00100 + G00101 2482 none all
L SmartLadder n/a n/a n/a n/a

On the gel it looks like lane 9, containing 013K, has the correct length. We will do a plasmid isolation tomorrow. Unfortunately none of the colonies of 020A seemed to be correct. We will check some more colonies with a colony PCR overnight.

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