IGEM:Cambridge/2008/Turing Pattern Formation/Primers: Difference between revisions
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We made primer sets for 2 ribosomal binding sites in Bacillus of differing binding strengths. We expect B._sub_RBSs to bind very strongly because it is the consensus sequence for RBS in B. subtilis. | We made primer sets for 2 ribosomal binding sites in Bacillus of differing binding strengths. We expect B._sub_RBSs to bind very strongly because it is the consensus sequence for RBS in B. subtilis. | ||
< | <pre> | ||
B. subtilis consensus RBS - http://www.ncbi.nlm.nih.gov/pubmed/10446248 | B. subtilis consensus RBS - http://www.ncbi.nlm.nih.gov/pubmed/10446248 | ||
| Line 9: | Line 9: | ||
B._sub_RBSs_R | B._sub_RBSs_R | ||
*5' CTGCAGCGGCCGCTACTAGTAACA CCTCCTTT CTCT 3'</ | *5' CTGCAGCGGCCGCTACTAGTAACA CCTCCTTT CTCT 3'</pre> | ||
< | <pre>RBS modified from ECE112 SpoVG RBS - http://www.bgsc.org/Catalogs/Catpart4.pdf | ||
B._sub_RBSw_F | B._sub_RBSw_F | ||
| Line 18: | Line 18: | ||
B._sub_RBSw_R | B._sub_RBSw_R | ||
*5' CTGCAGCGGCCGCTACTAGTAACA CCACCTCT CTCT 3'</ | *5' CTGCAGCGGCCGCTACTAGTAACA CCACCTCT CTCT 3'</pre> | ||
| Line 26: | Line 26: | ||
=====agr senders===== | =====agr senders===== | ||
< | <pre>Agr_B_F | ||
*5' GTTTCTT C GAATTC GCGGCCGC T TCTAG ATGaactattttgacaacaa 3' | *5' GTTTCTT C GAATTC GCGGCCGC T TCTAG ATGaactattttgacaacaa 3' | ||
| Line 36: | Line 36: | ||
Agr_D_R | Agr_D_R | ||
*5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA ttcatgcagctgggtcagct 3'</ | *5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA ttcatgcagctgggtcagct 3'</pre> | ||
=====agr receivers===== | =====agr receivers===== | ||
< | <pre>Agr_C_F | ||
*5' GTTTCTT C GAATTC GCGGCCGC T TCTAG atgattctgatgttcaccat 3' | *5' GTTTCTT C GAATTC GCGGCCGC T TCTAG atgattctgatgttcaccat 3' | ||
| Line 51: | Line 51: | ||
Agr_A_R | Agr_A_R | ||
*5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA gattttcttgacattgcgta 3' | *5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA gattttcttgacattgcgta 3' | ||
</ | </pre> | ||
Revision as of 09:37, 5 August 2008
Bacillus RBS Primers
We made primer sets for 2 ribosomal binding sites in Bacillus of differing binding strengths. We expect B._sub_RBSs to bind very strongly because it is the consensus sequence for RBS in B. subtilis.
B. subtilis consensus RBS - http://www.ncbi.nlm.nih.gov/pubmed/10446248 B._sub_RBSs_F *5' GAATTCGCGGCCGCTTCTAGAG AAAGGAGG TGTTA 3' B._sub_RBSs_R *5' CTGCAGCGGCCGCTACTAGTAACA CCTCCTTT CTCT 3'
RBS modified from ECE112 SpoVG RBS - http://www.bgsc.org/Catalogs/Catpart4.pdf B._sub_RBSw_F *5' GAATTCGCGGCCGCTTCTAGAG AGAGGTGG TGTTA 3' B._sub_RBSw_R *5' CTGCAGCGGCCGCTACTAGTAACA CCACCTCT CTCT 3'
Agr Primers
- These primers are to extract each part of the Agr operon so that a Bacillus RBS can replace the E. coli RBS currently attached to the genes.
agr senders
Agr_B_F *5' GTTTCTT C GAATTC GCGGCCGC T TCTAG ATGaactattttgacaacaa 3' Agr_B_R *5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA tttcagatcctctttgatg 3' Agr_D_F *5' CTT C GAATTC GCGGCCGC T TCTAG atgaatactctgttcaatctgtttt 3' Agr_D_R *5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA ttcatgcagctgggtcagct 3'
agr receivers
Agr_C_F *5' GTTTCTT C GAATTC GCGGCCGC T TCTAG atgattctgatgttcaccat 3' Agr_C_R *5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA gttattgatgatttcgactt 3' Agr_A_F *5' GTTTCTT C GAATTC GCGGCCGC T TCTAG atggaaatcgcactggcta 3' Agr_A_R *5' TT C CTGCAG CGGCCGC T ACTAGT A TTATTA gattttcttgacattgcgta 3'
