User:Torsten Waldminghaus/Primer

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  • All primers are in concentrations of 100pmol/μL
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Name Sequence Characteristics Primer number
GFPrv ctgatttaatctgtatcaggc The following primers are for the mutagenesis of GFP to introduce GATC sites as silent mutation (GFPrv, GFPfw, GFPmut1fw, GFPmut2rv, GFPmut3fw, GFPmut4rv, GFPmut5fw, GFPmut6fw, GFPmut7fw, GFPmut8fw, GFPmut9fw) Tm 55 (salt adjusted)[1] 1
GFPfw CTCTCTACTGTTTCTCCATAC Tm 57 2
GFPmut1fw caaacaaaagaatggGatcaaagctaac Tm 62 [without mutated nucleotide] 3
GFPmut2rv caatgttgtggcgGatCttgaagttag Tm 63 [without mutated nucleotide] 4
GFPmut3fw caactagcagaTcattatcaacaaaatac Tm 63 [without mutated nucleotide] 5
GFPmut4rv gccatcgccGatCggagtattttg Tm 62 [without mutated nucleotide] 6
GFPmut5fw cgaaaagcgtgaTcacatggtcc Tm 64 [without mutated nucleotide] 7
GFPmut6fw ctgctgctgggatCacacatggc Tm 66 [without mutated nucleotide] 8
GFPmut7fw GTTATCCGGACCATATGAAACGGC 5’-phosph. HPLC 9
GFPmut8fw CATTGAAGATGGGTCCGTTCAACTAG 5’-phosph. HPLC 10
GFPmut9fw CCTTTCGAAAGACCCCAACGAAAAG 5’-phosph. HPLC 11
GATC19DNAfw GCCCGCGGATCCGCCCGCC oligo for methylation experiment from A. Humeny et al. 2003 12
GATC19DNArv GGCGGGCGGATCCGCGGGC oligo for methylation experiment from A. Humeny et al. 2003 13
pyrBfw ggatcCACCCATTCCCAGCCCCTC cloning of pyrB 14
pyrBrv gaattcCCTTACAGTACCAGATCGCG cloning of pyrB 15
MseIlong AGTGGGATTCCGCATGCTAGT 16
MseIshort TAACTAGCATCG This sequence seems to be wrong because the last CG is not complementary to MseIlong but should instead be GC => use MseIshortnew instead 17
MseIshortnewNo TAACTAGCATGC Not phosphorilated 18
MseIshortnew TAACTAGCATGC 5'modified with phosphate 19
bet-fw GTCGACCCACAGGAACTGAT To distinguish DY330 (with lambda phage) from other E. coli strains use primers for bet as part of the recombination machinery of lambda 20
bet-rev GGCTGACGTTCTGCAGTGTA To distinguish DY330 (with lambda phage) from other E. coli strains use primers for bet as part of the recombination machinery of lambda 21
Cut-test_fw TAATAGGCATGCTAGCAGCTG AGCGTAGCTAGCGATGCAACGAC GGTGATCAGCGTCGATGCAGCCG AAACGATGACTGTCACATAGCTG ATGCTTTGCT
Cut_test_rv TAAGCAAAGCATCAGCTATGT GACAGTCATCGTTTCGGCTGCAT CGACGAAGATCACCGTCGTTGCA TCGCTAGCTACGCTCAGCTGCTA GCATGCCTAT
Cut-test_fw TAATAGGCATGCTAGCAGCTG AGCGTAGCTAGCGATGCAACGAC GGTGATCAGCGTCGATGCAGCCG AAACGATGACTGTCACATAGCTG ATGCTTTGCT
Cut_test_rv TAAGCAAAGCATCAGCTATGT GACAGTCATCGTTTCGGCTGCAT CGACGAAGATCACCGTCGTTGCA TCGCTAGCTACGCTCAGCTGCTA GCATGCCTAT
Cm_seqF
Cm_seqR