Kimberley and Jazz followed the protocol for the gel electrophoresis for the optimised G Luc, which showed presence of the gene. They then ordered more Tap media to grow algal cultures. Kimberley also contacted Carbrooke primary school to arrange a date for the SAW project, which was confirmed to be Wednesday 14th September. Ben looked through the biobrick library, to search for more biobricks and then later in the afternoon, he designed the algal plasmids. Gurdeep and Mark poured plates with antibiotic resistance for transforming the biobricks with E.coli. In the afternoon, Gurdeep helped design and constructed the presentation for the UK team meet up. Ben Hardy designed primers for use in the moss plasmid. Mario and Mark carried out biobrick duty in the afternoon, by searching for compatible biobricks, which lead to a grand total of 60, ready for tomorrows transformations!
Jazz and Kimberley completed a second mini prep for the optimised G Luc gene with a slightly modified protocol. Jazz then completed a restriction digestion, and ran a gel to check we have successfully isolated the plasmid. Kimberley finalised the lesson plan which was emailed out to those helping in the primary school along with the various worksheets. She also emailed out to try and get an alternative source of funding to support outreach activities. As of today the date for the next primary school was also confirmed. Jazz and Mario also measured the optical density of the algae to ensure we have a high enough cell density for transformations and plated out TAP media ready for tomorrow. Mario and Jazz made up algal cultures; two inoculated from the sample sent from chlammy centre and the third made from extracting 200 (microlitres) of our originally grown culture. Ben Hardy and Abbie selected suitable antibiotic resistance genes, designed their plasmid for moss, then completed primer designs and ordered them. Gurdeep, Mark, Ben Jevans and Mario carried out the 60 biobrick transformations into E.coli, starting at 9am and finishing at 5pm, tough day! Mark and Mario had to go and make new plates with antibiotic resistance as we were short by 15 tetracyclin and 6 ampicilin plates, so that we could carry on with the protocol. Now the plates are in the incubator, hopefully we will have colonies tomorrow!
Gurdeep, Ben Jevans , Mark and Mario made cultures of the transformed E.coli and left them to grow overnight. Gurdeep also helped to construct the presentation for the Friday meeting at the UEA and the UK meet up for the 25th July 2011. Kimberley, Jazz and Mario completed the algae transformation with the help of Ellis O’neil. Kimberley and Jazz attended a 2:00pm meeting with Kay Yeoman to go through the lab work the children would be doing and finalise arrangements for our first public engagement activity.
Gurdeep, Ben Jevans, Mark and Mario preformed glycerol stock to preserve the E.coli with the characteristic plasmids containing the 40 biobricks, followed by mini prep in order to isolate the plasmid. They then did a gel electrophoresis in order to confirm the presence of DNA, which took till 2pm, leading to a late lunch. After lunch Ben Jevans and Gurdeep worked on the new header and navigation bar for the new wiki. Kimberley finished the tree of life and printed off pictures for the synthetic biology workshop that her and Jazz successfully completed in the afternoon.
Gurdeep demonstrated how to edit and update the wiki in order for other members of the team to contribute to the wiki in the morning until lunch. Abbie and Ben Hardy made cultures of E.coli containing the CaMV promoter and Nos terminator, and left them to grow overnight. Ben, Kimberley and Jazz wrote a draft presentation which they presented to the whole team and a couple of advisors at UEA in preparation for the UK team meet on Monday.