Difference between revisions of "IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/19"

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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> Week Seven</span>
 
|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> Week Seven</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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Also transformations were done on the plasmids in order to make more of the DNA.
 
Also transformations were done on the plasmids in order to make more of the DNA.
  
The gel from the protein expression was imaged, it showed that sigma factor antA had been overexpressed in samples 1b and 2b (lanes 6 and 8) which had been induced by IPTG.
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The SDS-PAGE from the protein expression was imaged, it showed that sigma factor AntA had been overexpressed in samples 1b and 2b (lanes 6 and 8) which had been induced by IPTG.
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[[Image:Sds page jpeg.JPG|thumb|Figure 1: A 10% SDS-PAGE, stained with “Instant Blue”. Lanes 1-4 and 5-8 are insoluble and soluble fractions, respectively. Lanes 1, 3, 5 & 7 are samples without IPTG added; Lanes 2, 4, 6 & 8 are samples that had IPTG added.A broadrange SDS PAGE ladder was used]]
  
 
===Floor 1===
 
===Floor 1===
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Made 6 x 500 ml flasks of SFM, autoclaved and poured into plates to set (25 ml each).
 
Made 6 x 500 ml flasks of SFM, autoclaved and poured into plates to set (25 ml each).
  
Colony streaking - some plates need longer to grow as no Strep. has appeared or the colonies arent sporulating yet. Streaked all Strep.-like colonies and interesting looking colonies.
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Colony streaking - Streaked all <i>Streptomyces</i> single colonies and interesting looking colonies. Some plates need longer to grow for colonies to appear or they aren't sporulating yet.
  
 
==Outreach==
 
==Outreach==

Latest revision as of 23:05, 26 September 2017

Igem-logo-150px.png Week Seven Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Lab

Floor 2

Following the sequencing analysis from yesterday, we decided to do restriction enzyme digest reactions for our AntGp-Neo biobrick to fix the duplicated Xba1 sites, so that they get removed. Samples were run on a gel and fragments were cut and purified, DNA was stored in the freezer. Also transformations were done on the plasmids in order to make more of the DNA.

The SDS-PAGE from the protein expression was imaged, it showed that sigma factor AntA had been overexpressed in samples 1b and 2b (lanes 6 and 8) which had been induced by IPTG.

Figure 1: A 10% SDS-PAGE, stained with “Instant Blue”. Lanes 1-4 and 5-8 are insoluble and soluble fractions, respectively. Lanes 1, 3, 5 & 7 are samples without IPTG added; Lanes 2, 4, 6 & 8 are samples that had IPTG added.A broadrange SDS PAGE ladder was used

Floor 1

Numbered and labelled further samples collected (39-51).

Made 6 x 500 ml flasks of SFM, autoclaved and poured into plates to set (25 ml each).

Colony streaking - Streaked all Streptomyces single colonies and interesting looking colonies. Some plates need longer to grow for colonies to appear or they aren't sporulating yet.

Outreach

An excel sheet containing all the details about the soil and sediment samples was updated.