Molecular Cloning

From 2008.igem.org

(Difference between revisions)
(Template Resources)
(Primers)
Line 39: Line 39:
== Primers ==
== 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 ==
== PCR ==

Revision as of 13:55, 29 October 2008

HOME Team Project 1 Project 2 Parts Modelling Notebook Doodle Board

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

HOME Team Project 1 Project 2 Parts Modelling Notebook Doodle Board