Team:TU Delft/16 August 2010 content

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(Digestion)
(Alkane degradation)
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The digestions were executed for 15 minutes at 37 degrees, and checked on 1% agarose gel:
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The digestions were executed for 15 minutes at 37 degrees, and checked on [http://2010.igem.org/Team:TU_Delft#page=protocols/agarose_gel 1% agarose gel]:
[[Image:|300px|thumb|left|Digestion check]]
[[Image:|300px|thumb|left|Digestion check]]

Revision as of 16:26, 16 August 2010

Alkane degradation

Unfortunately last week's attempts hadn't yielded any BioBricks making use of the 2-way ligation method. We thus decided to take another stab at the 3-way ligation procedure, according to the BioBrick Assembly Manual of Ginkgo Bioworks. The BioBrick numbers we tried to construct were: 014C, 020C, 405C, 327C and 304C.

Digestion

# Sample Enzyme 1 Enzyme 2 Enzyme 3 Buffer BSA Needed fragment
1 011A EcoRI SpeI AseI NEBuffer 2 ‘E - J23100-J61100-alkB2-J61100-rubA3 - S’
2 013K XbaI PstI HindIII NEBuffer 2 ‘X - J61100-rubA4-J61100-rubR-B0015 - P’
3 017A EcoRI SpeI AseI NEBuffer 2 ‘E - J23100-J61100-ladA - S’
4 018A XbaI PstI AseI NEBuffer 2 ‘X - J61101-ADH - P’
5 402A EcoRI SpeI AseI NEBuffer 2 ‘E - J23100-J61101-PhPFDα - S’
6 403A XbaI PstI AseI NEBuffer 2 ‘X - J61101-PhPFDβ - P’
7 pCaiF EcoRI SpeI AseI NEBuffer 2 ‘E - pCaiF - S’
8 B0032 XbaI PstI AseI NEBuffer 2 ‘X - B0032 - P’
9 B0032 XbaI PstI AseI NEBuffer 2 ‘X - B0032 - P’
10 pAlkS EcoRI SpeI AseI NEBuffer 2 ‘E - pAlkS - P’
11 pSB1C3 EcoRI PstI None NEBuffer 2 ‘E - pSB1C3 - P’

The digestions were executed for 15 minutes at 37 degrees, and checked on 1% agarose gel:


[[Image:|300px|thumb|left|Digestion check]]