User:Torsten Waldminghaus/Primer

• 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		22
Cut_test_rv	TAAGCAAAGCATCAGCTATGT GACAGTCATCGTTTCGGCTGCAT CGACGAAGATCACCGTCGTTGCA TCGCTAGCTACGCTCAGCTGCTA GCATGCCTAT		23
Cut-test-meth_fw	TAATAGGCATGCTAGCAGCTG AGCGTAGCTAGCGATGCAACGAC GGTG'''A'''TCAGCGTCGATGCAGCCG AAACGATGACTGTCACATAGCTG ATGCTTTGCT	modified at position 49 with N6-methyl-dA	24
Cut_test-meth_rv	TAAGCAAAGCATCAGCTATGT GACAGTCATCGTTTCGGCTGCAT CGACGAAGATCACCGTCGTTGCA TCGCTAGCTACGCTCAGCTGCTA GCATGCCTAT	modified at position 53 with N6-methyl-dA	25
gatRfw	cgctttctcgaacaaaaagg		26
gatRrv	atgaatcgagaacggcaatc		27
yoeAfw	actgcaaccgcaacttcttc		28
yoeArv	cactacctcaacacgctcca		29
ter-check-fw	tcgtactggtgatggaacga	For checking the insertion of GATC cluster near terC with outside primers	30
ter-check-rv	aggattcacgcgataagtgg		31
T7fw	GAAATTAATACGACTCACTATAGGG		32
ORIrvT7	CACCGATCATTCACAGTTAATGATCCTTTCCAGGTTGTTGATCTTAAAAGCCGGATCCTTGCCCTATAGTGAGTCGTATTAATTTC		33
ORIfwT7	CAAGGATCCGGCTTTTAAGATCAACAACCTGGAAAGGATCATTAACTGTGAATGATCGGTGCCCTATAGTGAGTCGTATTAATTTC		34
ORIfwBIO	CAAGGATCCGGCTTTTAAGATCAACAACCTGGAAAGGATCATTAACTGTGAATGATCGGTG	5'-biotin labeled	35
ORIrv	CACCGATCATTCACAGTTAATGATCCTTTCCAGGTTGTTGATCTTAAAAGCCGGATCCTTG		36
Cm_seqF
Cm_seqR