Boston UniversityStatus

From 2007.igem.org

(Difference between revisions)
(Protocols)
(Short-Term To-Do List)
 
(61 intermediate revisions not shown)
Line 3: Line 3:
==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 ==
-
PrimersNeed to Buy
+
Error-Prone PCRFrom 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)
+
[[Boston_UniversityGel | Making an Electrophoresis Gel]]
-
Ordering of Error-Prone PCR Materials:  Not completed
+
[[Boston_UniversityZymo/SOB | Procedure for 50 mL E. Coli mixture in Zymo Broth or SOB
-
 
+
]]
-
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"]]
+
== 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]

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