Bcanton:Primers: Difference between revisions
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*C7077 - 51C & 57/58C | *C7077 - 51C & 57/58C | ||
*Gene 1 - 55C 58C | *Gene 1 - 55C 58C | ||
==12/6/05== | |||
'''Project''' - B0037 | |||
'''Forward homology sequence''' - 5'-aattcgcggccgcttctagaggtgtgtctagta-3' | |||
'''Reverse homology sequence''' - 5'-CTAGTACTAGACACACCTCTAGAAGCGGCCGCG-3' | |||
'''Reverse homology sequence reverse complement''' - 5'--3' | |||
'''5' additions''' already cut E/S | |||
'''Prefix''' - 5'--3' | |||
'''Suffix''' - 5'--3' | |||
'''Suffix reverse complement''' - 5'--3' | |||
'''Forward primer''' - 5'-aattcgcggccgcttctagaggtgtgtctagta-3' | |||
'''Reverse primer''' - 5'-CTAGTACTAGACACACCTCTAGAAGCGGCCGCG-3' | |||
'''Notes''' - | |||
'''Project''' - R8237 | |||
'''Forward homology sequence''' - 5'-aattcgcggccgcttctagagtaatacgtctcactatagggagatactagaggtgtgtctagtactagta-3' | |||
'''Reverse homology sequence''' - 5'-CTAGTACTAGACACACCTCTAGTATCTCCCTATAGTGAGACGTATTACTCTAGAAGCGGCCGCG-3' | |||
'''Reverse homology sequence reverse complement''' - 5'--3' | |||
'''5' additions''' already cut E/S | |||
'''Prefix''' - 5'--3' | |||
'''Suffix''' - 5'--3' | |||
'''Suffix reverse complement''' - 5'--3' | |||
'''Forward primer''' - 5'-aattcgcggccgcttctagagtaatacgtctcactatagggagatactagaggtgtgtctagtactagta-3' | |||
'''Reverse primer''' - 5'-CTAGTACTAGACACACCTCTAGTATCTCCCTATAGTGAGACGTATTACTCTAGAAGCGGCCGCG-3' | |||
'''Notes''' - | |||
'''Project''' - R4037 | |||
'''Forward homology sequence''' - 5'-aattcgcggccgcttctagagtccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagaggtgtgtctagtactagta-3' | |||
'''Reverse homology sequence''' - 5'-CTAGTACTAGACACACCTCTAGTAGTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGACTCTAGAAGCGGCCGCG-3' | |||
'''Reverse homology sequence reverse complement''' - 5'--3' | |||
'''5' additions''' | |||
'''Prefix''' - 5'--3' | |||
'''Suffix''' - 5'--3' | |||
'''Suffix reverse complement''' - 5'--3' | |||
'''Forward primer''' - 5'-aattcgcggccgcttctagagtccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagaggtgtgtctagtactagta-3' | |||
'''Reverse primer''' - 5'-CTAGTACTAGACACACCTCTAGTAGTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGACTCTAGAAGCGGCCGCG-3' | |||
'''Notes''' - |
Revision as of 13:29, 6 December 2005
Tools
Template
Project - Forward homology sequence - 5'--3' Reverse homology sequence - 5'--3' Reverse homology sequence reverse complement - 5'--3' 5' additions Prefix - 5'--3' Suffix - 5'--3' Suffix reverse complement - 5'--3' Forward primer - 5'--3' Reverse primer - 5'--3' Notes -
BioBrick prefixes and suffixes
- BioBrick prefix - 5'-GAATTC GCGGCCGC T TCTAGA G-3'
- BioBrick prefix for coding regions - 5'-GAATTC GCGGCCGC T TCTAG-3'
- BioBrick suffix - 5'-TACTAGT A GCGGCCG CTGCAG-3'
- BioBrick suffix reverse complement - 5'-CTGCAG CGGCCGC T ACTAGTA-3'
11/8/05
T7 gene 1 - To PCR T7 gene 1 from the T7 genome with BioBrick ends Forward homology sequence - ATG AAC ACG ATT AAC ATC GCT AAG AAC GAC TTC Reverse homology sequence - 5'-TTA CGC GAA CGC GAA GTC CG-3' I want to order it ready to cut X/P so that I can ligate downstream of a promoter-RBS Prefix - 5'-CGC T TCTAG-3' Suffix - 5'-TAC TAG TAG CGG CCG CTG CAG CTG G-3' Suffix reverse complement - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA-3' T71F - 5'-CGCTTCTAG ATG AAC ACG ATT AAC ATC GCT AAG AAC GAC TTC-3' TM1 - 60C TM2 - 63C T71R - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA TTA CGC GAA CGC GAA GTC CG-3' TM1 - 60C TM2 - 72C
C0011 - ccdB - Biobricking the ccdB gene from E. coli. Sequence from pDEST14(Invitrogen) courtesy of Heather Forward homology sequence - 5'-ATG CAG TTT AAG GTT TAC ACC TAT AAA AGA GAG -3' Tm - 57C Reverse homology sequence - 5'-CCA TTA ACC TGA TGT TCT GGG GAA TAT AA-3' Reverse homology sequence reverse complement - 5'-TTA TAT TCC CCA GAA CAT CAG GTT AAT GG-3' Tm - 56.5C 5' additions - Will be cutting X/P Prefix - 5'-CGC T TCTAG-3' Suffix - 5'-TAC TAG TAG CGG CCG CTG CAG CTG G-3' Suffix reverse complement - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA-3' C0011F - 5'-CGC T TCTAG ATG CAG TTT AAG GTT TAC ACC TAT AAA AGA GAG-3' Tm - 61.4C C0011R - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA TTA TAT TCC CCA GAA CAT CAG GTT AAT GG-3' Tm 69.1C Notes - Just the usual biobrick homodimers
T7 Gene 5.3 - C7053 - Forward homology sequence - 5'-ATG AAC GAA AGA CAC TTA ACA GGT GC-3' Tm = 57.6C Reverse homology sequence - 5'-GGC AAG AGA AAC AAA GGC ATA AAA CTA TAG-3' Reverse homology sequence reverse complement - 5'-CTA TAG TTT TAT GCC TTT GTT TCT CTT GCC -3' Tm - 56.4C 5' additions Prefix - 5'-CGC T TCTAG-3' Suffix - 5'-TAC TAG TAG CGG CCG CTG CAG CTG G-3' Suffix reverse complement - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA-3' T75.3F - 5'-CGC T TCTAG ATG AAC GAA AGA CAC TTA ACA GGT GC-3' Tm - 62.6C T75.3R - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA CTA TAG TTT TAT GCC TTT GTT TCT CTT GCC-3' Tm- 69.2C Notes -
T7 Gene 7.7 - C7077 - Forward homology sequence - 5'-GTG GAA GAC TGC ATT GAA TGG AC-3' Tm = 56.3C Reverse homology sequence - 5'-CAG CGT AAG ACA TGG AGA CAC ATT TA-3' Reverse homology sequence reverse complement - 5'-TAA ATG TGT CTC CAT GTC TTA CGC TG-3' Tm - 56.8C 5' additions Prefix - 5'-CGC T TCTAG-3' Suffix - 5'-TAC TAG TAG CGG CCG CTG CAG CTG G-3' Suffix reverse complement - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA-3' T77.7F - 5'-CGC T TCTAG GTG GAA GAC TGC ATT GAA TGG AC-3' Tm - 63.2C T77.7R - 5'-C CAG CTG CAG CGG CCG CTA CTA GTA TAA ATG TGT CTC CAT GTC TTA CGC TG-3' Tm - 69.7C Notes -
R2182 - Forward homology sequence - 5'-cgcttctagagtaatacgtctcactatagggagatactagagtcacacaggaaagtactagtagcggccgctgcagCTGG-3' Reverse homology sequence - 3'-CTTAAGCGCCGGCGAAGATCTCATTATGCAGAGTGATATCCCTCTATGATCTCAGTGTGTCCTTTCATGATCATCGCCGGCGACGTCGACC-5' Reverse homology sequence reverse complement - 5'-CCAGCTGCAGCGGCCGCTACTAGTACTTTCCTGTGTGACTCTAGTATCTCCCTATAGTGAGACGTATTACTCTAGAAGCGGCCGCGAATTC-3' 5' additions - Already ready to be cut X/P Prefix - 5'--3' Suffix - 5'--3' Suffix reverse complement - 5'--3' R2182F - 5'--3' R2182R - 5'--3' Notes -
R6182 - Forward homology sequence - 5'-gaattcgcggccgcttctagagtaatacgtctcactatagggagatactagaggtgtgtactagtagcggccgctgcagCTGG-3' Reverse homology sequence - 3'-CTTAAGCGCCGGCGAAGATCTCATTATGCAGAGTGATATCCCTCTATGATCTCCACACATGATCATCGCCGGCGACGTCGACC-5' Reverse homology sequence reverse complement - 5'-CCAGCTGCAGCGGCCGCTACTAGTACACACCTCTAGTATCTCCCTATAGTGAGACGTATTACTCTAGAAGCGGCCGCGAATTC-3' 5' additions - Already ready to be cut X/P Prefix - 5'--3' Suffix - 5'--3' Suffix reverse complement - 5'--3' R6182F - 5'--3' R6182R - 5'--3' Notes -
11/14/05
PCR conditions
- C0011 - 51C & 56C
- C7053 - 51C & 57C
- C7077 - 51C & 57/58C
- Gene 1 - 55C 58C
12/6/05
Project - B0037 Forward homology sequence - 5'-aattcgcggccgcttctagaggtgtgtctagta-3' Reverse homology sequence - 5'-CTAGTACTAGACACACCTCTAGAAGCGGCCGCG-3' Reverse homology sequence reverse complement - 5'--3' 5' additions already cut E/S Prefix - 5'--3' Suffix - 5'--3' Suffix reverse complement - 5'--3' Forward primer - 5'-aattcgcggccgcttctagaggtgtgtctagta-3' Reverse primer - 5'-CTAGTACTAGACACACCTCTAGAAGCGGCCGCG-3' Notes -
Project - R8237 Forward homology sequence - 5'-aattcgcggccgcttctagagtaatacgtctcactatagggagatactagaggtgtgtctagtactagta-3' Reverse homology sequence - 5'-CTAGTACTAGACACACCTCTAGTATCTCCCTATAGTGAGACGTATTACTCTAGAAGCGGCCGCG-3' Reverse homology sequence reverse complement - 5'--3' 5' additions already cut E/S Prefix - 5'--3' Suffix - 5'--3' Suffix reverse complement - 5'--3' Forward primer - 5'-aattcgcggccgcttctagagtaatacgtctcactatagggagatactagaggtgtgtctagtactagta-3' Reverse primer - 5'-CTAGTACTAGACACACCTCTAGTATCTCCCTATAGTGAGACGTATTACTCTAGAAGCGGCCGCG-3' Notes -
Project - R4037 Forward homology sequence - 5'-aattcgcggccgcttctagagtccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagaggtgtgtctagtactagta-3' Reverse homology sequence - 5'-CTAGTACTAGACACACCTCTAGTAGTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGACTCTAGAAGCGGCCGCG-3' Reverse homology sequence reverse complement - 5'--3' 5' additions Prefix - 5'--3' Suffix - 5'--3' Suffix reverse complement - 5'--3' Forward primer - 5'-aattcgcggccgcttctagagtccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagaggtgtgtctagtactagta-3' Reverse primer - 5'-CTAGTACTAGACACACCTCTAGTAGTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGACTCTAGAAGCGGCCGCG-3' Notes -