IGEM:Stanford/2009/Project Homeostasis/Helpful Websites
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Project |
Research Proposal |
Systems Overview |
Cloning Plan |
Sequences & Primers |
Anti-Inflammation |
Device Overview |
Parts Design |
Challenges |
Results |
Anti-Immunosuppression |
Device Overview |
Parts Design |
Challenges |
Results |
Protocols |
Modeling |
Overview |
Notebook |
Results |
Future Work |
Archived Work |
General Guidelines
1.18-30 nucleotide bases
2. Always end in G or C (for the stronger triple bond)
3. %GC should be 40-65%
4. Melting temp between 52-68C is good
5. Forward Primer:
- 1. Add TATA at the very beginning
- 2. After TATA add the proper prefix found on the registry (see below)
- 3. Check both initial and TATA prefix ratios and melting temps on IDT (see below)
6. Primer 2
- 1. Take end sequence of proper length then find the reverse complement (Ape see below)
- 2. Check the reverse complement for %GC and melting temp.
- 3. Add TATA and suffix to the beginning of the reverse complement strand (parts registry)
- 4. Check both initial and final sequence on IDT
7. Analyze both strands at NUPACK for hairpins and trouble spots
- 1. Insert both primer sequences and run the program
- 2. Shows potential areas for hairpin
- 3. Red = BAD
8.Both primers should be ready to go
9.Document all papers, gene sequences, and sites