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	[[Boston_UniversityPrimerDesign | Primer Design]]		[[Boston_UniversityPrimerDesign | Primer Design]]
-	== Week's (Ambitious) Goals ==	+	[[Boston_UniversityCompTransf | Making Shewanella Competent and Transforming Plasmid]]
-		+
-	Week of 6/4:	+
-		+
-	1. Evaluate the transformation that was done on Friday.	+
-		+
-	2. Confirm the correct plasmid (pJQ200)	+
-		+
-	3. Find appropriate restriction enzymes that cut by BLASTing all the plasmid's restriction enzyme sites onto the global transcription factors.	+
-		+
-	4. Re-design primers for the global transcription factors based on the restriction enzymes we have selected.	+
-		+
-	5. Order primers.	+
-		+
-	6. Practice regular (non-error prone) PCR with the primers to check that they work.	+
-		+
-	7. Incorporate the global transcription factors into the plasmid and transform this plasmid into the E.coli.	+
-		+
-	8. Conjugate this plasmid into Shewy.	+
	== Materials We Need ==		== Materials We Need ==
-	Error-Prone PCR:  Need to Buy	+	Error-Prone PCR:  From CAB(?)
-	Restriction Enzymes:  bought	+	Ligases:  Need to Buy
-	Ligases:  Need to Buy?	+	== Short-Term To-Do List ==
		+	EDIT THE WIIIIKIIII
-	TOPO kit:  available in lab	+	== Protocols ==
		+	[[Boston_UniversityHeatShockProtocol | Calcium Chloride/Heat Shock Plasmid Transformations Protocol]]
-	== Short-Term To-Do List ==	+	[[Boston_UniversityFilterConjugationProtocol | Filter Cojugation Protocol]]
-	Ordering of Error-Prone PCR Materials:  Not completed	+	[[Boston_UniversityTOPO | pTrcHis TOPO TA Expression Kit Cloning Protocol]]
-	Thank-You Letters sent to Pfizer:  Not Completed	+	[[Boston_UniversityNEBCutter| Using NEBCutter for checking specific restriction enzymes against a sequence]]
-	Thank-You Letters sent to BU ppl:  Not completed	+	[[Boston_UniversityGel | Making an Electrophoresis Gel]]
-	== Protocols ==	+	[[Boston_UniversityZymo/SOB | Procedure for 50 mL E. Coli mixture in Zymo Broth or SOB
-	[[Boston_UniversityHeatShockProtocol | "Calcium Chloride/Heat Shock Plasmid Transformations"]]	+	]]
	== Question and Answer ==		== Question and Answer ==
-	Can we get the kanR gene with appropriate sticky ends for BsaI and Tth111I? (DB)
-
-	== Relevant Publications and Links==
-	http://www.shewybase.bu.edu
-
-	https://www.atcc.org/common/catalog/numSearch/numResults.cfm?collection=mb-vector&atccNum=77482
	[https://2007.igem.org/Boston_University Back]		[https://2007.igem.org/Boston_University Back]

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Latest revision as of 16:43, 6 July 2007

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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

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