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	==What We've Accomplished==		==What We've Accomplished==
-	[[Boston_UniversityPrimerDesign | Primer Design]]	+	[[Boston_UniversityPlasmidChoice | Plasmid Choice]]
-	[[Boston_UniversityRestrictionEnzymePlasmid | Plasmid and Restriction Enzyme Choice]]	+	[[Boston_UniversityRestrictionChoice | Restriction Enzyme Choice]]
-	== Week's (Ambitious) Goals ==	+	[[Boston_UniversityPrimerDesign | Primer Design]]
-	Wednesday 5/30	+	[[Boston_UniversityCompTransf | Making Shewanella Competent and Transforming Plasmid]]
-	# Get all protocols	+
-	#  Identify materials/prepare order	+
-	#  Design Primers	+
-	#  Learn about budget/POs	+
-		+
-	Thursday 5/31	+
-	# Do primer order	+
-	# Start conjugation practice	+
-	# Confirm restriction enzymes, ligases	+
-	# Order confirmed/needed materials	+
-	# Team Revew Meeting	+
-	# Draft Thank-You Letters for our Sponsors	+
-		+
-	Friday 6/1	+
-	# Evaluate/continue conjugation, practice electroporation for E. coli	+
-	# Revise proposal to include possibility of screening with alginate beads and fluorocytometer	+
-	# Meeting with Tim:  Budgets/protocols, Pfizer/fundraising, iGEM registration, beads	+
-		+
-	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 ==
-	Primers:  Need to Buy	+	Error-Prone PCR:  From CAB(?)
-	Error-Prone PCR:  Need to Buy	+	Ligases:  Need to Buy
-		+
-	Plasmids:  Need to Buy?	+
-		+
-	Restriction Enzymes:  Need to Buy?	+
-		+
-	Ligases:  Need to Buy?	+
	== Short-Term To-Do List ==		== Short-Term To-Do List ==
		+	EDIT THE WIIIIKIIII
-	Lab Orientation: COMPLETED!	+	== Protocols ==
-	*[[Lab Orientation Checklist]]	+	[[Boston_UniversityHeatShockProtocol | Calcium Chloride/Heat Shock Plasmid Transformations Protocol]]
-	Lab Safety Training: COMPLETED!	+	[[Boston_UniversityFilterConjugationProtocol | Filter Cojugation Protocol]]
-	Design of Primers:  COMPLETED!	+	[[Boston_UniversityTOPO | pTrcHis TOPO TA Expression Kit Cloning Protocol]]
-	Ordering of Primers:  COMPLETED!	+	[[Boston_UniversityNEBCutter| Using NEBCutter for checking specific restriction enzymes against a sequence]]
-		+
-	Gathering of Protocols:  Not Completed (Chris, please send me the protocols when they are gathered)	+
-		+
-	Ordering of Error-Prone PCR Materials:  Not completed	+
-		+
-	Thank-You Letters sent to Pfizer:  Not Completed	+
-		+
-	Thank-You Letters sent to BU ppl:  Not completed	+
-		+
-	== Protocols ==	+
-	[[Boston_UniversityHeatShockProtocol | "Calcium Chloride/Heat Shock Plasmid Transformations"]]	+
		+	[[Boston_UniversityGel | Making an Electrophoresis Gel]]
		+	[[Boston_UniversityZymo/SOB | Procedure for 50 mL E. Coli mixture in Zymo Broth or SOB
		+	]]
	== Question and Answer ==		== Question and Answer ==
-
-	== Relevant Publications and Links==
-	http://www.shewybase.bu.edu
	[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