NYMU Taipei/Help me/planning
From 2007.igem.org
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<p> </p> | <p> </p> | ||
<h3>Primer design</h3> | <h3>Primer design</h3> | ||
| - | <p>RcsC uses | + | <p>RcsC uses [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&qty=1&c_start=1&list_uids=48994873&uids=&dopt=gbwithparts&dispmax=5&sendto=&fmt_mask=0&from=2315049&to=2318000&extrafeatpresent=1&ef_CDD=8&ef_MGC=16&ef_HPRD=32&ef_STS=64&ef_tRNA=128&ef_microRNA=256&ef_Exon=512#sequence_48994873 UUG] as start codon!!</p> |
<p>Location of transmembrane signal</p> | <p>Location of transmembrane signal</p> | ||
<p>[http://openwetware.org/wiki/Synthetic_Biology:BioBricks/Part_fabrication#Quick_reference BioBrick standard]</p> | <p>[http://openwetware.org/wiki/Synthetic_Biology:BioBricks/Part_fabrication#Quick_reference BioBrick standard]</p> | ||
Revision as of 14:42, 18 October 2007
Phase 1
- 把含50px的plasmid transform E. coli，拿到plasmid DNA。（需要：protocol, reagent）
- 把tar-Envz中的Envz切下（RE site: NdeI, ?)，接到cloning vector上（需要：protocol, 實驗室有的vector）<a href="http://partsregistry.org/Part:BBa_M0040">LVA tag</a>
- 把RcsC用PCR分離出來 (內含AflII)（Reverse primer RE site須設計: NdeI），接到cloning vector上 (需要把每個domain的DNA sequence搞清楚)
- 設計primer時須考慮ribosome binding site的問題
需要人力；每一任務需要一人全力投入。
C3CCBKRB016
Primer design
RcsC uses UUG as start codon!!
Location of transmembrane signal
IGFBP3
IGFBP7
RcsC:
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 1 21 55.92 38.10 4.00 2.00 GAAATACCTTGCTTCTTTTCG
RIGHT PRIMER 1417 20 58.14 45.00 8.00 2.00 GAACATCGATTTTGACTGGC
Phase 2
- 把RcsC和Envz接在一起。
- 把Reporter接到OmpR binding site後面。
- Quantitative characterization of promotor efficiency (basal transcription level)
RcsC primer:
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 1 21 55.92 38.10 4.00 2.00 GAAATACCTTGCTTCTTTTCG
RIGHT PRIMER 1417 20 58.14 45.00 8.00 2.00 GAACATCGATTTTGACTGGC
