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Contents 1 What We've Accomplished 2 Materials We Need 3 Short-Term To-Do List 4 Protocols 5 Question and Answer

What We've Accomplished

Plasmid Choice

Restriction Enzyme Choice

Primer Design

Making Shewanella Competent and Transforming Plasmid

Materials We Need

Error-Prone PCR: From CAB(?)

Ligases: Need to Buy

Short-Term To-Do List

David: 1)Look up metal-sensitive/redox-sensitive chelating dyes for Tim's grant 2)Look up how to control alginate bead size for Tim's grant 3)Begin primer design for pJQ200 (regulator, promoter, global transcription regulators) 4)send stuff to Danny for wiki update 5)grow overnight of the shewie zymo for miniprep tomorrow

Rahul: 1)Work on compiling documents, protocols, results and send to Danny for Wiki updates 2)Understand error-prone PCR and write a short summary/outline of important details and key steps (and problem areas and controls)

Christian: 1)Decide on promoters, regulators to put into pJQ200 2)document this stuff so we can wiki it

Danny: 1)Wiki needs a format update /new look. we'll email some suggestions for you to look at later today 2)receive info from us and update wiki

Protocols

Calcium Chloride/Heat Shock Plasmid Transformations Protocol

Filter Cojugation Protocol

pTrcHis TOPO TA Expression Kit Cloning Protocol

Using NEBCutter for checking specific restriction enzymes against a sequence

Making an Electrophoresis Gel

Procedure for 50 mL E. Coli mixture in Zymo Broth or SOB

Question and Answer

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