Wiki/Team:Warsaw/primers

From 2008.igem.org

(Difference between revisions)
Line 54: Line 54:
<h4>Mutagenesis of the A protein</h4>
<h4>Mutagenesis of the A protein</h4>
<pre>
<pre>
-
<a name="AmutDelecjaL+KpnI">AmutDelecjaL+KpnI</a>   5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
+
<a name="AmutDelecjaL+KpnI">AmutDelecjaL+KpnI</a> 5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
-
<a name="AmutDelecjaP">AmutDelecjaP</a>   5' AACTTAAACGAAGAACAACGAAACGCC 3'
+
<a name="AmutDelecjaP">AmutDelecjaP</a>     5' AACTTAAACGAAGAACAACGAAACGCC  3'
-
<a name="AmutMniejL+KpnI">AmutMniejL+KpnI</a>  5' AGTTAGGTACCGGTGAGATCTCATAGGACGCGTTTTGTTGTTCTTTGTTG  3'
+
<a name="AmutWiecejL+KpnI">AmutWiecejL+KpnI</a> 5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
-
<a name="AmutMniejP">AmutMniejP</a>  5' TAAACGAAGAACAAGGAAACGCCTTCATCCAAAGTTTAAAAGATGACCC 3'
+
<a name="AmutWiecejP">AmutWiecejP</a>       5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'
-
<a name="AmutWiecejL+KpnI">AmutWiecejL+KpnI</a>   5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
+
-
<a name="AmutWiecejP">AmutWiecejP</a>   5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'
+
</pre>
</pre>
<h4>Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein</h4>
<h4>Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein</h4>
<pre>  
<pre>  
-
<a name="AL+link10+homo2">AL+link10+homo2</a>   5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
+
<a name="AL+link10+homo2">AL+link10+homo2</a>     5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
-
<a name="AlphaP+link10+homo2">AlphaP+link10+homo2</a>   5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
+
<a name="AlphaP+link10+homo2">AlphaP+link10+homo2</a> 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
-
<a name="OmegaP+link10+homo2">OmegaP+link10+homo2</a>   5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'
+
<a name="OmegaP+link10+homo2">OmegaP+link10+homo2</a> 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'
-
<a name="AL+SacI">AL+SacI</a>   5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
+
<a name="AL+SacI">AL+SacI</a>             5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
-
<a name="AP+NotI">AP+NotI</a>   5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
+
<a name="AP+NotI">AP+NotI</a>             5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
-
<a name="AlphaL+SacI">AlphaL+SacI</a>   5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
+
<a name="AlphaL+SacI">AlphaL+SacI</a>         5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
-
<a name="OmegaL+SacI">OmegaL+SacI</a>   5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
+
<a name="OmegaL+SacI">OmegaL+SacI</a>         5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
-
<a name="AlphaPlinkSac">AlphaPlinkSac</a>   5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'
+
<a name="AlphaPlinkSac">AlphaPlinkSac</a>       5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'
</pre>
</pre>
<br><br><br><br>
<br><br><br><br>
Line 90: Line 88:
<a name="PETt7L_XNE">pETt7L_XNE</a>          5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3'
<a name="PETt7L_XNE">pETt7L_XNE</a>          5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3'
<a name="PlacL_XNE">PlacL_XNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3'
<a name="PlacL_XNE">PlacL_XNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3'
-
<a name="AP+XhoI+SpeI">AP+XhoI+SpeI</a>   5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'
+
<a name="AP+XhoI+SpeI">AP+XhoI+SpeI</a>         5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'
</pre>
</pre>
</tr></table>
</tr></table>
</html>
</html>

Revision as of 15:50, 26 September 2008

Gallery Bricks Notebook Team Project Home

 

Primers

One-armed bandit preparation

AID and T7 cloning

T7lLinkB   5' CGGATCCGGCGGTGGCGGTAGTGGAGGAGGTGGGTCTGGAGGCGGAGGGAACACGATTAACATCGCTAAGAAC  3'
T7lRBSHi   5' CACAAGCTTAGGAGGTGTACTAGATGAACACGATTAACATCGCTAAG  3'
T7pXbSal   5' TCTGTCGACTCTAGATTACGCGAACGCGAAGTCC  3'
AIDlNrH    5' CGTAAGCTTAGGAGGTGTAGACCATGGACAGCCTCTTGATG  3'
AIDpLinB   5' CCACTACCGCCACCGCCGGATCCGCCACCTCCGCTGCCTCCACCACCAAGTCCCAAAGTACGAAATGC  3'

Sequencing of pMPM-T5

pMPMT5seqF  5' CTACTGTTTCTCCATACCCG  3'
pMPMT5seqR  5' GAAAATCTTCTCTCATCCGC 3'
AIDseqP     5' GCTTCGGCGCATCCTTTTG 3'

Sequencing of β-galactosidase from pZC320

pZCseqL     5' CTTGACCCGCAGTTGCAAAC 3'
pZCseqP     5' CCCAACTGATCTTCAGCATC 3'

Sequencing of the T7 polymerase gene

T7Rseq      5' GCTTCAACCGCAGCTACCAC 3'

Hunter and prey

Fusion of protein A and ompA and alpha or omega part of beta-lactamase

alphaL+link14+homo (alphaL_link)          5' CAGTGGTGGAAGCGGTGGC GGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGT CACCCAGAAACGCTGGTGAAAG 3'
alphaP+STOP+XbaI+BamHI (alphaP_XB)        5' GG GGATCC GG TCTAGA TTA TTCGCCAGTTAATAGTTTGCG 3'
omegaL+link14+homo (omegaL_link)          5' CAGTGGTGGAAGCGGTGGC GGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGT CTACTTACTCTAGCTTCCCGG 3'
omegaP+STOP+EcoRI+PstI+BamHI (omegaP_EPB) 5' AAGGATCCAACTGCAGAAGAATTC TTACCAATGCTTAATCAGTG 3'
OmpAL+NdeI (ompaL_N)                      5' CATATGAAAGCTACTAAACTGGTACTGGG 3'
OmpAP+link8+SacI+NotI+homo (ompaP_link)   5' GCCACCGCTTCCACCACT GCGGCCGC CA GAGCTC CCTCCTCCAGAACCTCC GCCGTTGTCCGGACGAGTGCCGATG 3'

Cloning of A and/or Z protein to expression vector (pET15b/pET24a)

AlphaP+link+Sac   5' GG GAGCTC CC ACCACCCCCACCCCCGCTACCTC  3'
AlphaL+Nde        5' CGGG CATATG CACCCAGAAACGCTGGTGAAAG  3'

Mutagenesis of the A protein

AmutDelecjaL+KpnI 5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
AmutDelecjaP      5' AACTTAAACGAAGAACAACGAAACGCC  3'
AmutWiecejL+KpnI  5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
AmutWiecejP       5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'

Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein

 
AL+link10+homo2     5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
AlphaP+link10+homo2 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
OmegaP+link10+homo2 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'
AL+SacI             5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
AP+NotI             5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
AlphaL+SacI         5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
OmegaL+SacI         5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
AlphaPlinkSac       5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'




BioBricks Preparation

AIDP_HindSpeNotPst   5' AAACTGCAGCGGCCGCTACTAGTAAAGCTTTCAAAGTCCCAAAGTACGAAATG  3'
AL_BNXNE             5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
AlfaPSpe             5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'
APSacSpe             5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3'
LinLSXNE             5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC  3'
LinP_BS              5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC  3'
LinPBSNP             5' AGCCTGCAGCGGCCGCTACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTACC  3'
OmegPSpe             5' GCACTAGTATTACCAATGCTTAATCAGTGAG  3'
OmegLNde             5' CAGCCATATGCTACTTACTCTAGCTTCCCGG  3'
OmpLNXNE             5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
OmLNXNEB             5' ATTAGATCTGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
pETt7L_XNE           5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3'
PlacL_XNE            5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3'
AP+XhoI+SpeI         5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'

Retrieved from "http://2008.igem.org/Wiki/Team:Warsaw/primers"