Team:TU Delft/5 August 2010 content

From 2010.igem.org

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(→Emulsifier)
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==Alkane degradation==
==Alkane degradation==
 +
===colony PCR===
The morning started with a gel checking the PCR products of the one BioBrick that didn't give positives [https://2010.igem.org/Team:TU_Delft#page=pages/blog&blog=5_August_2010 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.
The morning started with a gel checking the PCR products of the one BioBrick that didn't give positives [https://2010.igem.org/Team:TU_Delft#page=pages/blog&blog=5_August_2010 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.
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|6
|6
|rest
|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:
 +
 +
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
 +
|'''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
|}
|}

Revision as of 15:47, 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.

TUDelft groningen DSC5932.JPG TUDelft groningen DSC5934.JPG

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.

TU Delft Pi4 2010-08-04 Colony PCR.png

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.

PCR of 020 colonies

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
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