Team:TU Delft/6 August 2010 content
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Latest revision as of 10:37, 13 October 2010
Contents |
Lab work
Alkane degration
Yesterday's plates showed colonies. To check whether these contained the desired insert a colony PCR was performed. The products of this PCR were checked on a gel.
On the gel, two colonies of 011A showed the right length. None of the lengths for 013A or K were correct. They did however match the PCR length (009A, 482) of one the inserts which was digested in order to create this BioBrick. Next week we will take a few more colonies from the plate and check these, and otherwise the digestion and ligation will be tried again.
Lane description
# | Description | Primers | Expected length (bp) | ✓ | ✗ |
L | SmartLadder | n/a | n/a | n/a | n/a |
1-5 | 011A | G00100 + G00101 | 1816 | 1,5 | 2,3,4 |
6-10 | 013A | G00100 + G00101 | 1929 | none | 6,7,8,9,10 |
11-15 | 013K | G00100 + G00101 | 1929 | none | 11,12,13,14,15 |
L | SmartLadder | n/a | n/a | n/a | n/a |
Hydrocarbon Sensing
Ligation
Digested pAlkS, B0032, AlkS and P(Caif) were ligated as followed:
# | Description | Volume (µl) |
1 | PalkS12-B0032 | 9.5 |
AlkS-E0240 | 3.0 | |
ligase | 1.5 | |
ligase buffer | 1 | |
2 | PalkS12-B0032 | 10.5 |
pSB1T3 | 2.0 | |
ligase | 1.5 | |
ligase buffer | 1 | |
3 | P(Caif)-B0032 | 9.5 |
AlkS-E0240 | 3.0 | |
ligase | 1.5 | |
ligase buffer | 1 | |
4 | P(Caif)-B0032 | 10.5 |
pSB1T3 | 3.0 | |
ligase | 1.5 | |
ligase buffer | 1 |
The ligation products were checked on a gel.