Team:York UK/Notebook.html

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iGEM York

Future in Your Eyes, future in York UNI team iGEM York Team
Future in Your Eyes, future in York UNI team

Overview

My part of the team - Gintare, Evaldas and Jonas - were responsible for cloning of the mtr complex, gold sensing device and preparing peptides for expression.

July 7th

Making of chemically competent DH5α cells. Protocol can be found in our protocols section.

July 10th

Purification of genomic DNA from Shewanella oneidensis and Salmonella sp.

July 25th

After a long break we finally started working on the actual assembly of parts - the mtr complex. Today Jonas ran a trial PCR on mtrB and mtrC part of the complex both of which are around the size of 2kb. GoTaq polymerase (Promega) was used, this is the polymerase we generally use for trials and colony PCRs as fidelity is not the main priority.

mtrBCtrialThis is the gel from the PCR. Temperatures ranged 46-71˚C. It appears that the primers are sufficiently specific at even low temperatures. 1% agarose, SYBR-Safe(Invitrogen), QStep4 ladder


July 26th

Trial PCR again, this time for three small (~500bp) synthetic DNA fragments - gBlocks (IDT), these fragments are highlighted below on the diagram as well as the overall view of our mtr gene. mtrABC

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University of York

The University of York

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