Team:TU Delft/5 August 2010 content
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[[Image:TU_Delft_Pi4_2010-08-04_Colony_PCR.png|400px]] | [[Image:TU_Delft_Pi4_2010-08-04_Colony_PCR.png|400px]] | ||
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+ | Lane Description: | ||
+ | {|style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" | ||
+ | |'''#''' | ||
+ | |'''Description''' | ||
+ | |'''Expected Length (bp)''' | ||
+ | |'''Primers''' | ||
+ | |'''Status''' | ||
+ | |'''Remarks''' | ||
+ | |- | ||
+ | |1 | ||
+ | |SmartLadder | ||
+ | |n/a | ||
+ | |n/a | ||
+ | |n/a | ||
+ | | | ||
+ | |- | ||
+ | |2 | ||
+ | |BBa_K398101 | ||
+ | |929 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=green>✓</font> | ||
+ | | | ||
+ | |- | ||
+ | |3 | ||
+ | |BBa_K398101 | ||
+ | |929 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |4 | ||
+ | |BBa_K398101 | ||
+ | |929 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |5 | ||
+ | |BBa_K398101 | ||
+ | |929 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |6 | ||
+ | |BBa_K398101 | ||
+ | |929 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |7 | ||
+ | |BBa_K398402 | ||
+ | |758 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |8 | ||
+ | |BBa_K398402 | ||
+ | |758 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |9 | ||
+ | |BBa_K398402 | ||
+ | |758 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=green>✓</font> | ||
+ | | | ||
+ | |- | ||
+ | |10 | ||
+ | |SmartLadder | ||
+ | |n/a | ||
+ | |n/a | ||
+ | |n/a | ||
+ | | | ||
+ | |- | ||
+ | |11 | ||
+ | |BBa_K398402 | ||
+ | |758 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |- | ||
+ | |12 | ||
+ | |BBa_K398402 | ||
+ | |758 | ||
+ | |G00101 + G00100 | ||
+ | |<font color=red>✗</font> | ||
+ | | | ||
+ | |} | ||
==Alkane degradation== | ==Alkane degradation== |
Revision as of 15:48, 5 August 2010
Contents |
Groningen Team
Today three members had an appointment with one of the professors at our department. We heard of this and invited them to come over to meet each other.
Lab Work
Alkane Sensing, Solvent Tolerance and Salt Tolerance
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.
Lane Description:
# | Description | Expected Length (bp) | Primers | Status | Remarks |
1 | SmartLadder | n/a | n/a | n/a | |
2 | BBa_K398101 | 929 | G00101 + G00100 | ✓ | |
3 | BBa_K398101 | 929 | G00101 + G00100 | ✗ | |
4 | BBa_K398101 | 929 | G00101 + G00100 | ✗ | |
5 | BBa_K398101 | 929 | G00101 + G00100 | ✗ | |
6 | BBa_K398101 | 929 | G00101 + G00100 | ✗ | |
7 | BBa_K398402 | 758 | G00101 + G00100 | ✗ | |
8 | BBa_K398402 | 758 | G00101 + G00100 | ✗ | |
9 | BBa_K398402 | 758 | G00101 + G00100 | ✓ | |
10 | SmartLadder | n/a | n/a | n/a | |
11 | BBa_K398402 | 758 | G00101 + G00100 | ✗ | |
12 | BBa_K398402 | 758 | G00101 + G00100 | ✗ |
Alkane degradation
colony PCR
The morning started with a gel checking the PCR products of the one BioBrick that didn't give positives yesterday. This gel showed one positive, but when I centrifuged the cells, they gave a red pellet. This indicated that the plasmid unfortunately was not the desired BioBrick, but the original destination plasmid.
Lane description
# | Description | Primers | Expected length (bp) | ✓ | ✗ |
L | SmartLadder | n/a | n/a | n/a | n/a |
1-9 | 020K | G00100 + G00101 | 2428 | 6 | rest |
Plasmid Isolation
Of the colonies which gave a positive PCR, the plasmids were isolated using a QIAgen miniprep kit. The follwoing concentrations were obtained:
The following plasmid concentrations were obtained:
BioBrick | Composed of | Concentration (ng/μL) |
007A (1) | J61100-alkB2 | 33.8 |
007A (2) | J61100-alkB2 | 32.0 |
012K | J61100-rubR-B0015 | 42.7 |
021A | J61107-ALDH-B0015 | 46.3 |
021K (5) | J61107-ALDH-B0015 | 7.3 |
021K (1) | J61107-ALDH-B0015 | 11.1 |
021K (2) | J61107-ALDH-B0015 | 12.9 |