Boston UniversityStatus
From 2007.igem.org
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== Question and Answer == | == Question and Answer == | ||
| + | According to ATCC, plasmid PJQ200 "replicates only in enterobacteria, and so functions as a suicide vector in other Gram negative hosts." Since shewy is not an enterobacterium, how will PJQ200 help us get that gene into shewy and its future generations? | ||
== Relevant Publications and Links== | == Relevant Publications and Links== | ||
Revision as of 13:32, 8 June 2007
Contents |
What We've Accomplished
Week's (Ambitious) Goals
Wednesday 5/30
- 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
Primers: BOUGHT!
Error-Prone PCR: Need to Buy
Plasmids: BOUGHT!
Restriction Enzymes: Need to Buy?
Ligases: Need to Buy?
Short-Term To-Do List
Lab Orientation: COMPLETED!
Lab Safety Training: COMPLETED!
Design of Primers: COMPLETED!
Ordering of Primers: COMPLETED!
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
"Calcium Chloride/Heat Shock Plasmid Transformations"
Question and Answer
According to ATCC, plasmid PJQ200 "replicates only in enterobacteria, and so functions as a suicide vector in other Gram negative hosts." Since shewy is not an enterobacterium, how will PJQ200 help us get that gene into shewy and its future generations?
