IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/23

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(Floor Two)
Current revision (13:29, 22 August 2013) (view source)
(Floor Two)
 
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==Floor One==
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Made up 250 ml of LB Broth and 7000 ml of SFM + agar. All media was autoclaved for 1 hr and poured into plates to set.
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Diluted samples 45-59 (from 10^-1 to 10^-8).
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Set up cultures of Top10 and ETpuz E. coli cells transformed with pMS82 and pAU3-45. Added 10 ml LB (-NaCl) to 300 ul of cells and 10 ul of respective antibiotics - Hygromycin and Chloramphenicol for pMS82 and Apramycin and Chloramphenicol for pAU3-45.
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==Floor Two==
==Floor Two==
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The samples from the previous days minipreps were digested with the restriction enzyme Xba1 and the results analysed using Gel electrophoresis.
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Yesterday's minipreps were tested by (i) checking their concentrations (ii) performing restriction digestion reactions with Xba1 and analysing them using gel electrophoresis ''fig 1''.
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After discussing the results, samples 2b2 and 3b1 were selected for further experiments.
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[[Image:BIORAD 2013-07-23 16hr 38min.JPG|thumb|Fig.Analysis of Restriction digest products using gel electrophoresis. Each sample was cut with Xba1 and as a control the uncut DNA was loaded next to it. Even numbered lanes contain cut DNA and odd lanes contain uncut controls]]
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[[Image:23 july.JPG|thumb|Fig 1:Analysis of Restriction digest products using gel electrophoresis. Each sample was cut with Xba1 and as a control the uncut DNA was loaded next to it. Even numbered lanes contain cut DNA and odd lanes contain uncut controls]]
==Outreach==
==Outreach==

Current revision

Week 8 Main project page
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Floor One

Made up 250 ml of LB Broth and 7000 ml of SFM + agar. All media was autoclaved for 1 hr and poured into plates to set.

Diluted samples 45-59 (from 10^-1 to 10^-8).

Set up cultures of Top10 and ETpuz E. coli cells transformed with pMS82 and pAU3-45. Added 10 ml LB (-NaCl) to 300 ul of cells and 10 ul of respective antibiotics - Hygromycin and Chloramphenicol for pMS82 and Apramycin and Chloramphenicol for pAU3-45.

Floor Two

Yesterday's minipreps were tested by (i) checking their concentrations (ii) performing restriction digestion reactions with Xba1 and analysing them using gel electrophoresis fig 1.

After discussing the results, samples 2b2 and 3b1 were selected for further experiments.

Fig 1:Analysis of Restriction digest products using gel electrophoresis. Each sample was cut with Xba1 and as a control the uncut DNA was loaded next to it. Even numbered lanes contain cut DNA and odd lanes contain uncut controls
Fig 1:Analysis of Restriction digest products using gel electrophoresis. Each sample was cut with Xba1 and as a control the uncut DNA was loaded next to it. Even numbered lanes contain cut DNA and odd lanes contain uncut controls

Outreach

Today we had a visit from Ben Thompson, senior public relations officer at the Society of Microbiology. He spoke to us about our project and iGEM in general. The interview would be available as a podcast for the general public at a later date.

Figure 1: The Team being interviewed by Ben Thompson
Figure 1: The Team being interviewed by Ben Thompson



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