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 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 - 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.