Molecular Cloning

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(Difference between revisions)
(Primers)
(Primers)
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PP-PhaP-NotI-for
PP-PhaP-NotI-for
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<span style="color: Pink">5- TATGCGGCCGCTGTTTGTGCATTGCACAAAATCCA-3</span>
+
<span style="color: Brown">5- TATGCGGCCGCTGTTTGTGCATTGCACAAAATCCA-3</span>
PP-PhaP-HindIII-rev
PP-PhaP-HindIII-rev
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IGEM-ZouYLP-CreT7ter-rev-in
IGEM-ZouYLP-CreT7ter-rev-in
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<span style="color: Pink">5-AGCGGGGTTTTTTTTTCTACTAATCGCCATCTTCCAGCAG-3</span>
+
<span style="color: Brown">5-AGCGGGGTTTTTTTTTCTACTAATCGCCATCTTCCAGCAG-3</span>
IGEM-ZouYLP-CreT7ter-EcoRI-rev-out
IGEM-ZouYLP-CreT7ter-EcoRI-rev-out

Revision as of 14:02, 29 October 2008

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Molecular Cloning

Contents

Plasmids

   1 pUCPhbCAB

   2 pACYC(lacI+) DuetSRRz

   3 pACYC(lacI+) DuetEGFP

   4 pACYC(lacI-) DuetSRRz

   5 pACYC(lacI-) DuetEGFP

   6 pACYC(LacI-)DuetSRRzPpPhaPLacILVACret7terPrPhaRLoxPLacILoxP

   7 pACYC(LacI-)DuetEGFPPpPhaPLacILVACret7terPrPhaRLoxPLacILoxP

Template Resources

   PhbCAB: PBHR68 Plasmid
   Lac I: PET28a
   CRE:
   PpPhaP: Ralstonia eutropha H16 genome
   PrPhaR: Ralstonia eutropha H16 genome
   SRRz:
   EGFP:

Primers

(1) SRRz Primers

SRRZ-NcoI-for

5- ATCCATGGATGAAGATGCCAGAAAAACATGACCTGTTG-3

SRRZ-rev-in

5 - ACCCCGCCGAAGCGGGGTTTTTTTTTCTACTATCTGCACTGCTCATTAATA-3

SRRZ-BamHI-rev-out

5 -TATGGATCCAAAAAAAAACCCCGCCGAAGCGGGGTT-3

(2) Pp Operon primers

PP-PhaP-NotI-for

5- TATGCGGCCGCTGTTTGTGCATTGCACAAAATCCA-3

PP-PhaP-HindIII-rev

5- CACCATGTCGACTTTCTCCTCTTTAAGCTTTCAGGCAGCCGTCGTCTTCTTTG-3

LacILVA-HindIII-for

5-TGCCTGAAAGCTTAAAGAGGAGAAAGTCGACATGGTGAATGTGAAACCAGTAAC-3

LacILVA-rev-in

5- AGCTACTAAAGCGTAGTTTTCGTCGTTTGCAGCCTGCCCGCTTTCCAGTCGGGAAACCT-3

LacILVA-rev-PstI-out

5- ATTGGACATGCGCGCTTTCTCCTCTTTCTGCAGTTATTAAGCTACTAAAGCGTAGTTTT-3

IGEM-ZouYLP-CreT7ter—PstI-for

5- TGCAGAAAGAGGAGAAAGCGCGCATGTCCAATTTACTGACCGTACACCAAAATT-3

IGEM-ZouYLP-CreT7ter-rev-in

5-AGCGGGGTTTTTTTTTCTACTAATCGCCATCTTCCAGCAG-3

IGEM-ZouYLP-CreT7ter-EcoRI-rev-out

5-ATGAATTCGAGCTCGAAAAAAAAACCCCGCCGAAGCGGGGTTTTTTTTTCTACTAAT-3

(3) Pr operon primers

PR-PhaR-NotI-for

5-ATGCGGCCGCAGTGCCTTGTTGGGCATAGAATCAGGGCAGCGGCGCAGC-3

PR-PhaR-NdeI-rev

5-ACGAAGTTATCATATGTTATTACTTCTTGTCCGGCTGGTTGAACGGGAACGTCCCGAAC-3

LRLacILP-for-in

5-TGCTATACGAAGTTATAAAGAGGAGAAACTCGAGATGGTGAATGTGAAACCAGTAACGT-3

LRLacILP-NdeI-for-out

5-ACAAGAAGTAATAACATATGATAACTTCGTATAATGTATGCTATACGAAGTTATAAAGA-3

LRLacILP-rev-in

5-TACGAAGTTATCTACTGCCCGCTTTCCAGTCGGGAAACCT-3

LRLacILP-KpnI-rev-out

5-ATGGTACCATAACTTCGTATAGCATACATTATACGAAGTTATCTACTGCCCGCTT-3

(4) EGFP Primers

EGFP-for-KoZg-for

5-ccatgggcagcaagggcgaggagctgttc-3

EGFP-rev-in

5-AAAAAAAAACCCCGCCGAAGCGGGGTTTTTTTTTCTATCACTTGTACAGCTCGTC-3

EGFP-rev-out

5-TTTTC GAGCTC GAATTC GGATCCAAAAAAAAACCCCGCCGAA-3

PCR

Fusion PCR

Cut and Insert

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