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

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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 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.

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.
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.

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 - 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.

Outreach

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



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