Team:EPF-Lausanne/Notebook
From 2009.igem.org
(Difference between revisions)
Line 20: | Line 20: | ||
<br> 1.Promoter T7, RBS, CBP and LOVTAP: | <br> 1.Promoter T7, RBS, CBP and LOVTAP: | ||
:gtttcttcgaattcgcggccgcttctagagtaatacgactcactataggggaattgtg | :gtttcttcgaattcgcggccgcttctagagtaatacgactcactataggggaattgtg | ||
- | + | 2.RBS, CBP and LOVTAP: | |
:gtttcttcgaattcgcggccgcttctagagtgtttaactttaagaaggag | :gtttcttcgaattcgcggccgcttctagagtgtttaactttaagaaggag | ||
- | + | 3.CBP and LOVTAP: | |
:gtttcttcgaattcgcggccgcttctagatgaagcgacgatggaaaaagaatttcatag | :gtttcttcgaattcgcggccgcttctagatgaagcgacgatggaaaaagaatttcatag | ||
- | + | 4.LOVTAP: | |
:gtttcttcgaattcgcggccgcttctagatgctactacacttgaacgtattgagaagaac | :gtttcttcgaattcgcggccgcttctagatgctactacacttgaacgtattgagaagaac | ||
- | + | One reverse primer were designed: | |
:gtttcttcctgcagcggccgctactagtatcaatcgcttttcagcaacacctcttc | :gtttcttcctgcagcggccgctactagtatcaatcgcttttcagcaacacctcttc | ||
Revision as of 16:07, 6 July 2009
Contents |
Notebook
06.07.09
- Wet lab
LOVTAP plasmid AND TrpR plasmid were transformed in competent E. Coli, and grown overnight.
One problem: we actually don't know TrpR plasmid resistance, so we tried with three resistances available in the lab: Amp., Kana. and Chl.
LOVTAP is in a plasmid called pCal-n (see picture below):
Some comments on the plasmid:
-CBP is a small peptide with which we could purify LOVTAP protein
-Thrombin target is a nucleotidic sequence that can be recognized by thrombin a peptidase. This peptidase will cut CBP once LOVTAP is purified
- Cloning strategy
Four forward primers were designed to amplify:
1.Promoter T7, RBS, CBP and LOVTAP:
- gtttcttcgaattcgcggccgcttctagagtaatacgactcactataggggaattgtg
2.RBS, CBP and LOVTAP:
- gtttcttcgaattcgcggccgcttctagagtgtttaactttaagaaggag
3.CBP and LOVTAP:
- gtttcttcgaattcgcggccgcttctagatgaagcgacgatggaaaaagaatttcatag
4.LOVTAP:
- gtttcttcgaattcgcggccgcttctagatgctactacacttgaacgtattgagaagaac
One reverse primer were designed:
- gtttcttcctgcagcggccgctactagtatcaatcgcttttcagcaacacctcttc
Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook | Lectures | Team Management | References |
---|