IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/19: Difference between revisions
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===Floor 2=== | ===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. | 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. | |||
[[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=== | |||
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 <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== | ||
Revision as of 05:41, 25 September 2013
 Week Seven
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LabFloor 2Following 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.  Floor 1Numbered 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. OutreachAn excel sheet containing all the details about the soil and sediment samples was updated.
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