Team:TU Delft/2 August 2010 content

From 2010.igem.org

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Results of the digestion on [[Team:TU_Delft/protocols/agarose_gel|1% agarose gel]] 
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[[Image:TUDelft_20100802_digestion.png|400px|1% Agarose of digestion check ]]
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gel runned at 100V for 1 hour
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loaded 5 μL of marker and 10 μL sample + 2 μL loadingbuffer
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Lane description:
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{|style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
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|'''#'''
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|'''Description'''
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|'''Expected Length (bp)'''
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|'''Status'''
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|''Remarks'''
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|marker
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|1
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|Undigested part name + vector (e.g. B0015 in pSB1AK3)
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|✓or ✗
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|-
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|2
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|Part name + vector (e.g. B0015 in pSB1AK3) + Enzyme 1 + Enzyme 2
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|
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|✓or ✗
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|
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|-
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|3
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|Undigested part
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|✓or ✗
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|4
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|Part + Enzyme 1 + Enzyme 2
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|✓or ✗
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|5
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|Undigested part
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|✓or ✗
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|6
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|Part + Enzyme 1 + Enzyme 2
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|✓or ✗
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|7
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|Undigested part
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|✓or ✗
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|8
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|Part + Enzyme 1 + Enzyme 2
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|✓or ✗
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|9
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|Undigested part
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|✓or ✗
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|10
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|Part + Enzyme 1 + Enzyme 2
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|✓or ✗
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|-
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|11
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|Undigested part
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|✓or ✗
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|-
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|12
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|Part + Enzyme 1 + Enzyme 2
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|
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|✓or ✗
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|13
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|Undigested part
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|✓or ✗
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|14
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|Part + Enzyme 1 + Enzyme 2
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|
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|✓or ✗
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|
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|-
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|15
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|Undigested part
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|✓or ✗
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|-
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|16
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|Part + Enzyme 1 + Enzyme 2
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|✓or ✗
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|-
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|17
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|Undigested part
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|✓or ✗
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|-
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|18
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|Part + Enzyme 1 + Enzyme 2
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|✓or ✗
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|-
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|19
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|Undigested part
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|✓or ✗
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|-
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|20
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|Part + Enzyme 1 + Enzyme 2
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|
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|✓or ✗
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|
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Revision as of 09:29, 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:

# 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’

Results of the digestion on 1% agarose gel 1% Agarose of digestion check gel runned at 100V for 1 hour loaded 5 μL of marker and 10 μL sample + 2 μL loadingbuffer

Lane description:

# Description Expected Length (bp) Status Remarks'
marker
1 Undigested part name + vector (e.g. B0015 in pSB1AK3) ✓or ✗
2 Part name + vector (e.g. B0015 in pSB1AK3) + Enzyme 1 + Enzyme 2 ✓or ✗
3 Undigested part ✓or ✗
4 Part + Enzyme 1 + Enzyme 2 ✓or ✗
5 Undigested part ✓or ✗
6 Part + Enzyme 1 + Enzyme 2 ✓or ✗
7 Undigested part ✓or ✗
8 Part + Enzyme 1 + Enzyme 2 ✓or ✗
9 Undigested part ✓or ✗
10 Part + Enzyme 1 + Enzyme 2 ✓or ✗
11 Undigested part ✓or ✗
12 Part + Enzyme 1 + Enzyme 2 ✓or ✗
13 Undigested part ✓or ✗
14 Part + Enzyme 1 + Enzyme 2 ✓or ✗
15 Undigested part ✓or ✗
16 Part + Enzyme 1 + Enzyme 2 ✓or ✗
17 Undigested part ✓or ✗
18 Part + Enzyme 1 + Enzyme 2 ✓or ✗
19 Undigested part ✓or ✗
20 Part + Enzyme 1 + Enzyme 2 ✓or ✗
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