Team:Cambridge/Lab book/Week 13

From 2012.igem.org

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(Tuesday (18/09/12))
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Cultures induced with 1mM iptg or 3mM arabinose for luxBB, imaged after 3 hrs. Some extremely faint luminescence for 33,34,38 on photon counting camera.
Cultures induced with 1mM iptg or 3mM arabinose for luxBB, imaged after 3 hrs. Some extremely faint luminescence for 33,34,38 on photon counting camera.
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Cultures miniprepped and restriction digests carried out w/spa1. Results are shown. Digests are of 33-38, each miniprep, cut and uncut, between 2 ladders. Results are inconclusive.
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Cultures miniprepped and restriction digests carried out w/spa1. Results are shown. Results indicate problem with the miniprep and/or damage to the construct - expected bands at 6625, 2838, 1876 and 53 bp.
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[[File:salI.jpg]]
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[[File:salI.jpg|250px|right|thumb|Digests are of 33-38, each miniprep, cut and uncut, between 2 ladders.]]
===Wednesday (19/09/12)===
===Wednesday (19/09/12)===

Revision as of 14:33, 20 September 2012

Week: 3 4 5 6 7 8 9 10 11 12 13 14

Contents

Monday (17/09/12)

Testing of the 2.0 construct:

Numbers 33,35,36 have correct sequence as of being sent to us. However they were not visibly luminescent. *34 was.

Overnight cultures of each of the 6 culture lines sent were set up. Additionally, inducing plates were streaked, including the 2010 lux biobrick as a luminescence control and pjs130 as an iptg induction control.

Primers ordered to ligate 2.0 construct into pjs130 for insertion into bacillus.

Tuesday (18/09/12)

None of restreaked plates visibly luminescent, aside from *34 (faint), even under photon counting camera. Pjs130 not green, so iptg induction potentially didn't work. LuxBB induced and luminescent.

Cultures induced with 1mM iptg or 3mM arabinose for luxBB, imaged after 3 hrs. Some extremely faint luminescence for 33,34,38 on photon counting camera.

Cultures miniprepped and restriction digests carried out w/spa1. Results are shown. Results indicate problem with the miniprep and/or damage to the construct - expected bands at 6625, 2838, 1876 and 53 bp.

Digests are of 33-38, each miniprep, cut and uncut, between 2 ladders.

Wednesday (19/09/12)

Digest with different enzyme, hindIII, carried out. 15ul template DNA used in a 20ul reaction. Again, results were odd. Cut and uncut minipreps ran.

Cultures of ratiometrica were miniprepped to be transformed into bacillus. A restriction digest was performed on these minipreps, and produced bands of the correct size.

Thursday (20/09/12)

Friday (21/09/12)

Saturday (22/09/12)

Sunday (23/09/12)