Team:UEA-JIC Norwich/Weekeight

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Team meeting discussing finance and outreach. Alistair and Ben Hardy grew up bacteria containing a rbs biobrick. Ben Jevans and Gurdeep did a PCR. Kimberley started looking through images for the SAW project and played with ideas for a scientific activity which she could carry out in a school.

Ben Jevans and Gurdeep performed another site directed mutagenesis whilst Abbie, Ben Hardy and Alistair performed a double restriction digest of their plasmid to extract the desired promoter. Mario and Mark made new plates for our future transformations in algae.

Abbie, Ben Hardy and Alistair ran their digest on a gel and got successful bands. Next Abbie, Ben Hardy and Alistair performed a gel extraction to confirm and isolate their promoter followed by another nanodrop to find out the overall DNA concentration that was present. Finally Abbie, Ben Hardy and Alistair performed another digest to cut out their desired terminator from the plasmid.

Ben Jevans cultured the pSP1215 bacterial mutants in two LB broth liquid mediums containing the antibiotic phleomycin and two LB broth liquid mediums containing the antibiotic ampicilin. Abbie, Ben Hardy and Alistair administrated a gel electrophoresis of their digested Nos terminator. To further clarify their results they ran another gel which comprised of the whole plasmid running in collaboration with the digested promoter and terminator to ensure absolute clarity. All gels ran this day showed vivid bands and that all the experiments that had been performed this day had gone accordingly.

Ben Jevans observed his cultures he’d left to grow from the previous day; results showed that the cultures containing the antibiotic phleomycin had exhibited growth whilst those cultures containing ampicilin had no growth. Mario and Mark performed a nanodrop on previously mini-prepped G-Luc plasmids to identify which vial contained the highest DNA concentration. Mario and Mark then carried out a restriction digest to make the DNA more linearized and thus more well suited for transformation in algae. Mario and Mark then carried out another optical density count to identify we had an efficient amount of algal cells needed for a transformation. A transformation in chlamydomonas with yet more revisions to the original protocol for a more efficient transformation frequency was conducted. Abbie, Ben Hardy and Alistair performed another double restriction digest to attempt extracting the Nos terminator with a higher yield of DNA this time. Ben Jevans and Gurdeep also performed a mini-prep of their site-directed mutagenesis product constructed on the 2nd of august and then did a nanodrop.

Abbie, Ben Hardy and Alistair came in and did a PCR on the isolated CaMV promoter to try and make it into an applicable biobrick. They then ran it on a gel but got inconclusive results. Due to this Abbie, Ben Hardy and Alistair performed a miniprep to extract their transformed plasmid again. Gurdeep and Ben Jevans also came in and performed a miniprep and gel electrophoresis. However they did not observe any bands on the gel even though having a high concentration of DNA observed in the nano-drop.