Boston UniversityStatus

From 2007.igem.org

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[[Boston_UniversityPrimerDesign | Primer Design]]
[[Boston_UniversityPrimerDesign | Primer Design]]
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== Week's (Ambitious) Goals ==
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[[Boston_UniversityCompTransf | Making Shewanella Competent and Transforming Plasmid]]
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Week of 6/4:
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1. Evaluate the transformation that was done on Friday.
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2. Confirm the correct plasmid (pJQ200)
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3. Find appropriate restriction enzymes that cut by BLASTing all the plasmid's restriction enzyme sites onto the global transcription factors.
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4. Re-design primers for the global transcription factors based on the restriction enzymes we have selected.
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5. Order primers.
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6. Practice regular (non-error prone) PCR with the primers to check that they work.
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7. Incorporate the global transcription factors into the plasmid and transform this plasmid into the E.coli.
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8. Conjugate this plasmid into Shewy.
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== Materials We Need ==
== Materials We Need ==
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PrimersBOUGHT!
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Error-Prone PCRFrom CAB(?)
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Error-Prone PCR:  Need to Buy
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Ligases:  Need to Buy
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Plasmids:  BOUGHT!
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== Short-Term To-Do List ==
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EDIT THE WIIIIKIIII
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Restriction Enzymes:  Need to Buy?
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== Protocols ==
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[[Boston_UniversityHeatShockProtocol | Calcium Chloride/Heat Shock Plasmid Transformations Protocol]]
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Ligases:  Need to Buy?
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[[Boston_UniversityFilterConjugationProtocol | Filter Cojugation Protocol]]
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== Short-Term To-Do List ==
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[[Boston_UniversityTOPO | pTrcHis TOPO TA Expression Kit Cloning Protocol]]
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Ordering of Error-Prone PCR Materials:  Not completed
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[[Boston_UniversityNEBCutter| Using NEBCutter for checking specific restriction enzymes against a sequence]]
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Thank-You Letters sent to Pfizer:  Not Completed
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[[Boston_UniversityGel | Making an Electrophoresis Gel]]
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Thank-You Letters sent to BU ppl:  Not completed
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[[Boston_UniversityZymo/SOB | Procedure for 50 mL E. Coli mixture in Zymo Broth or SOB
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== Protocols ==
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[[Boston_UniversityHeatShockProtocol | "Calcium Chloride/Heat Shock Plasmid Transformations"]]
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== Question and Answer ==
== Question and Answer ==
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Can we get the kanR gene with appropriate sticky ends for BsaI and Tth111I? (DB)
 
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== Relevant Publications and Links==
 
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http://www.shewybase.bu.edu
 
[https://2007.igem.org/Boston_University Back]
[https://2007.igem.org/Boston_University Back]

Latest revision as of 16:43, 6 July 2007

Back

Contents

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

EDIT THE WIIIIKIIII

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

Back