Team:UEA-JIC Norwich/Weekeleven

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A nanodrop was carried out by Ben Jevans in the morning on the original miniprepped PCR G-Luc products from last week, results look outstanding. Three specific samples were chosen for a double restriction digestion and then the outcome of each digestion was ran on a gel; results on the gel look faint but evidently present. Ben Jevans then streak plated pSP124S E.coli samples on three plates slightly varying in their content, one agar plate contained ampicillin and the other two agar plates contained phleomycin. Kimberley gathered all the plates streaked with the colonies grown with the G-Luc plasmid, unfortunately the length of time it took to see if we had any transformed colonies caused contamination of the plates. The plates were discarded. Kimberley also made liquid cultures of transformed algae with linearized G-Luc.

Kimberley and Ben performed the new transformation technique electroporation in algae to try and attain better luck with getting successful transformants. This time also starch embedding was carried out to help increase the success rate of getting transformants and prevent the algae solution from drying out and dying on the plates. Ben Jevans also made cultures from his streak plates he had grown previously and performed a PCR to amplify the pSAD and heat-shock protein 70 promoters. The products of the PCR were then ran on another gel, in doing so we acquired more questionable results. Alistair and Abbie performed another moss transformation with water to test whether the transformation protocol worked.

Alistair grew up E.coli cultures from the RFP plate to be minipreped on Thursday 25th August.

Today Ben Jevans tests his pSP124S transformed cells grown in cultures containing different antibiotics. Firstly he mini-preps them and finally runs them on a gel; results yet again seem to be kind of promising but not definitive. Alistair digested the RFP plasmid and a random biobrick plasmid with xbaI and speI in a double digest, along with using each enzyme in a single digest.

Another PCR performed to amplify the pSAD terminator; when ran on the gel it certainly looks like something is there.