Team:Newcastle/Labwork/10 September 2009

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Three colonies were extracted from the 200ul ''cotC-GFP-smtA'' transformant cells plate and used, under aseptic conditions, to inoculate 3 x 5ml LB+kanamycin tubes. In the same way three colonies from the 200ul ''kinA'' transformant cells plate was used to inoculate 3 x 5ml LB+kanamycin tubes. These six tubes were then placed in the orbital shaking incubator at 37C for overnight growth.
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====Preparations for Midi-preps====
====Preparations for Midi-preps====
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Revision as of 23:18, 13 September 2009

Lab Work - 10/09/09

Metal Sensing Team

Introduction

Yesterday saw the Metal Sensing team attempt to transform DH5-alpha E.coli bacterial cells with two BioBricks (the cotC-GFP-smtA BioBrick and the kinA BioBrick) and plate them out on LB + kanamycin plates. The cotC-GFP-smtA BioBrick is part of the Metal Sensing team's sub-project and the kinA BioBrick will be used by the Stochastic Switch team's sub-project.

Today's exercise should see the Metal Sensing team inoculate LB medium with any transformant cultures which grow on these plates - if there are no cultures present then the transformations will be attempted a second time.

Observations

On all four plates there could be seen hundreds of transformant colonies - this was to be expected as the DNA used to transform the DH5-alpha cells was in pure form.

Procedure

There were two procedures carried out in response to these observations:

  • Three colonies from each 200ul plate of transformants were used to inoculate 6 x 5ml LB (making a total of six tubes of inoculated 5ml LB). This is preparation for mini-preps.


  • One colony from each of the 200ul plates (1 cotC-GFP-smtA transformed E.coli colony and 1 kinA transformed E.coli colony) was taken to inoculate 2 x 50ml LB media. This is preparation for midi-preps.


Preparations for Mini-preps

Three colonies were extracted from the 200ul cotC-GFP-smtA transformant cells plate and used, under aseptic conditions, to inoculate 3 x 5ml LB+kanamycin tubes. In the same way three colonies from the 200ul kinA transformant cells plate was used to inoculate 3 x 5ml LB+kanamycin tubes. These six tubes were then placed in the orbital shaking incubator at 37C for overnight growth.

Preparations for Midi-preps



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