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| - | {{EPF-Lausanne09}}
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| - | <div CLASS="epfltrick">__TOC__
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| - | </div><div CLASS="epfl09">
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| - |
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| - | =Cloning strategy=
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| - | ==July==
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| - |
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| - | ===06.07.09===
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| - | Four forward primers were designed to amplify:
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| - | <br> 1.Promoter T7, RBS, CBP and LOVTAP:
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| - | :gtttcttcgaattcgcggccgcttctagagtaatacgactcactataggggaattgtg
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| - | 2.RBS, CBP and LOVTAP:
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| - | :gtttcttcgaattcgcggccgcttctagagtgtttaactttaagaaggag
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| - | 3.CBP and LOVTAP:
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| - | :gtttcttcgaattcgcggccgcttctagatgaagcgacgatggaaaaagaatttcatag
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| - | 4.LOVTAP:
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| - | :gtttcttcgaattcgcggccgcttctagatgctactacacttgaacgtattgagaagaac
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| - | One reverse primer were designed:
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| - | :gtttcttcctgcagcggccgctactagtatcaatcgcttttcagcaacacctcttc
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| - |
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| - |
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| - | The '''recipient IGEM part''' have been chosen: [http://partsregistry.org/partsdb/get_part.cgi?part=BBa_B0010 BBa_B0010], well 13D in the received kit plate 1
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| - |
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| - | ===07.07.09===
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| - | To design plasmids : software Vector NTI
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| - |
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| - | ===08.07.09===
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| - | Inducible LOVTAP biobrick strategy
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| - |
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| - | ===09.07.09===
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| - | Partial digestion strategy.
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| - |
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| - | ===10.07.09===
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| - |
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| - | ===13.07.09===
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| - | Restriction enzymes on [http://www.neb.com/nebecomm/products/category1.asp?#2 Biolabs website]
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| - | and [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_olignucleotides.asp clevage oligonucleotides]
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| - |
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| - | TRP promoter biobrick strategy
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| - |
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| - | ===14.07.09===
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| - | Primers designed for LOVTAP read-out and RBphP project:
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| - |
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| - | 1.Forward primer Trp promoter:
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| - |
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| - | gtttcttc gaattcgcggccgcttctagagtggcaaatattctgaaatgagctgttgacaattaatcatcgaactagttaactagtacgc
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| - |
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| - | 2.Reverse primer Trp promoter:
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| - |
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| - | ctagctagctaggtcgataccctttttacgtgaacttgcgtactagttaactagttcgatgattaattgtca
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| - |
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| - | 3.1st Forward primer Inverter TetR:
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| - |
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| - | aatcatcgaactagttaactagtacgcaagttcacgtaaaaagggtatcgacaaagaggagaaatactagatgtcc
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| - |
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| - | 4.2nd Forward primer Inverter TetR:
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| - |
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| - | gtttcttcgaattcgcggccgcttctagagtggcaaatattctgaaatgagctgttgacaattaatcatcgaactagttaactagta
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| - |
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| - | 5.Reverse Primer Inverter TetR:
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| - |
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| - | ctagctagctag tttctcctctttctctagtagtgc
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| - |
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| - |
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| - | 6.Forward primer ppsR1 R.Palustris CGA009:
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| - |
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| - | gtttcttc*gaattcgcggccgcttctag*atgctggaggatatttgccctggtg
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| - |
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| - | 7.Reverse primer ppsR1 R.Palustris CGA009:
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| - |
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| - | gtttcttc*ctgcagcggccgctactagta*ttactcatcggctccgtctccttc
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| - |
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| - | 8.Forward primer ppsR2 R.Palustris CGA009:
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| - |
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| - | gtttcttc*gaattcgcggccgcttctag*atggcgtcaaagtccgttcatgcc
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| - |
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| - | 9.Reverse primer ppsR2 R.Palustris CGA009:
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| - |
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| - | gtttcttc*ctgcagcggccgctactagta*tcaatcctctgcgtcgtctgagg
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| - |
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| - |
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| - | 10.Forward primer BrBphP Bradyrhizobium ORS278:
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| - |
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| - | gtttcttc*gaattcgcggccgcttctag*atgcccgttccgctgacgac
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| - |
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| - | 11.Reverse primer BrBphP Bradyrhizobium ORS278:
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| - |
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| - | gtttcttc*ctgcagcggccgctactagta*tcactcctcgctctgcgagc
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| - |
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| - | 12.Forward primer ppsR1 Bradyrhizobium ORS278:
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| - |
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| - | gtttcttc*gaattcgcggccgcttctag*atgagggcgttcagagctcc
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| - |
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| - | 13.Reverse primer ppsR1 Bradyrhizobium ORS278:
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| - |
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| - | gtttcttc*ctgcagcggccgctactagta*ctattccaactgactgtcttcttcgc
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| - |
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| - | 14.Forward primer ppsR2 Bradyrhizobium ORS278:
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| - |
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| - | gtttcttc*gaattcgcggccgcttctag*atggccgagtttcacggtccac
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| - |
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| - | 15.Reverse primer ppsR2 Bradyrhizobium ORS278:
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| - |
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| - | gtttcttc*ctgcagcggccgctactagta*ctagctccccttttcggtttcctc
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| - |
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| - | ===15.07.09===
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| - |
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| - | ===16.07.09===
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| - |
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| - | ===17.07.09===
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| - |
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| - | ==August==
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| - |
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| - | </div><div CLASS="epfl09bouchon"></div>
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