Home  Contacts  Order  Catalog  Support
 Search  Alphabetical Index  Numerical Index
Fermentas logo
 Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification
 Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents
S U P P O R T
 

Digestion of Methylated DNA:

Effect of EcoKI and EcoBI Methylation on DNA Cleavage by Restriction Enzymes

Methylated DNA substrates were purified from E.coli K12 or E.coli B.

Enzymes which Recognition Sequences may Overlap EcoBI Methylase Site TGA(N)8TGCT
Single letter code:
R = G or A; 
Y = C or T; 
W = A or T; 
M = A or C; 		 K = G or T; 
S = C or G;
H = A, C or T;
V = A, C or G;		 B = C, G or T;
D = A, G or T;
N = G, A, T or C.

A = N6-methyladenine (m6A)
Recognition sequence is indicated in bold.

Enzyme Sequence Effect
AatII 5'...TGACGTC(N)4TGCT...3'
3'...ACTGCAG(N)4ACGA...5'		 not blocked
AluI 5'...TGACGTC(N)5TGCT...3'
3'...ACTGCAG(N)5ACGA...5'		 blocked
BclI
FastDigest® BclI		 5'...TGATCA(N)5TGCT...3'
3'...ACTAGT(N)5ACGA...5'		 blocked
BglII
FastDigest® BglII		 5'...TGAGATCT(N)3TGCT...3'
3'...ACTCTAGA(N)3ACGA...5'		 impaired
Bpu10I 5'...CCTGAGC(N)6TGCT...3'
3'...GGACTCG(N)6ACGA...5'		 not blocked
Bsp143I 5'...TGATC(N)6TGCT...3'
3'...ACTAG(N)6ACGA...5'		 not blocked
EcoRI
FastDigest® EcoRI		 5'...TGAATTC(N)4TGCT...3'
3'...ACTTAAG(N)4ACGA...5'		 not blocked
HincII
FastDigest® HincII		 5'...GTTGAC(N)7TGCT...3'
3'...CAACTG(N)7ACGA...5'		 blocked
HindIII
FastDigest® HindIII		 5'...TGAAGCTT(N)3TGCT...3'
3'...ACTTCGAA(N)3ACGA...5'		 impaired
HinfI
FastDigest® HinfI		 5'...TGANTC(N)5TGCT...3'
3'...ACTNAG(N)5ACGA...5'		 blocked
HphI 5'...GGTGA(N)8TGCT...3'
3'...CCACT(N)8ACGA...5'		 blocked
MboI
FastDigest® MboI		 5'...TGATC(N)6TGCT...3'
3'...ACTAG(N)6ACGA...5'		 blocked
MboII
FastDigest® MboII		 5'...TGAAGA(N)5TGCT...3'
3'...ACTTCT(N)5ACGA...5'		 not blocked
MluI
FastDigest® MluI		 5'...TGACGCGT(N)3TGCT...3'
3'...ACTGCGCA(N)3ACGA...5'		 not blocked
MnlI
FastDigest® MnlI		 5'...TGAGG(N)6TGCT...3'
3'...ACTCC(N)6ACGA...5'		 blocked
NdeI
FastDigest® NdeI		 5'...TGA(N)4CATATGCT...3'
3'...ACT(N)4GTATACGA...5'		 not blocked
OliI 5'...TGA(N)4CACGTGCT...3'
3'...ACT(N)4GTGCACGA...5'		 blocked
PaeI
FastDigest® SphI (PaeI)		 5'...TGA(N)5GCATGCT...3'
3'...ACT(N)5CGTACGA...5'		 blocked
PagI
FastDigest® BspHI (PagI)		 5'...TCATGA(N)8TGCT...3'
3'...AGTACT(N)8ACGA...5'		 impaired
SacI
FastDigest® SacI		 5'...TGAGCTC(N)4TGCT...3'
3'...ACTCGAG(N)4ACGA...5'		 not blocked
ScaI 5'...TGAGTACT(N)3TGCT...3'
3'...ACTCATGA(N)3ACGA...5'		 blocked
SspI
FastDigest® SspI		 5'...TGAATATT(N)3TGCT...3'
3'...ACTTATAA(N)3ACGA...5'		 not blocked
TasI 5'...TGAATT(N)5TGCT...3'
3'...ACTTAA(N)5ACGA...5'		 blocked

Enzymes which Recognition Sequences may Overlap EcoKI Methylase Site AAC(N)6GTGC
Single letter code:
R = G or A; 
Y = C or T; 
W = A or T; 
M = A or C; 		 K = G or T; 
S = C or G;
H = A, C or T;
V = A, C or G;		 B = C, G or T;
D = A, G or T;
N = G, A, T or C.

A = N6-methyladenine (m6A)
Recognition sequence is indicated in bold.

Enzyme Sequence Effect
Alw21I
FastDigest® Alw21I		 5'...AAC(N)6GTGCWC...3"
3'...TTG(N)6CACGWG...5'		 not blocked
Alw44I
FastDigest® ApaLI (Alw44I)		 5'...AAC(N)6GTGCAC...3'
3'...TTG(N)6CACGTG...5'		 not blocked
BseSI
FastDigest® Bme1580I (BseSI)		 5'...AAC(N)6GTGCMC...3'
3'...TTG(N)6CACGKG...5'		 impaired
DraI
FastDigest® DraI		 5'...TTTAAAC(N)6GTGC...3'
3'...AAATTTG(N)6CACG...5'		 blocked
HincII
FastDigest® HincII		 5'...GTYAAC(N)6GTGC...3'
3'...CARTTG(N)6CACG...5'		 blocked
KspAI
FastDigest® HpaI (KspAI)		 5'...GTTAAC(N)6GTGC...3'
3'...CAATTG(N)6CACG...5'		 blocked
MluI
FastDigest® MluI		 5'...AACGCGT(N)2GTGC...3'
3'...TTGCGCA(N)2CACG...5'		 not blocked
MssI
FastDigest® MssI		 5'...GTTTAAAC(N)6GTGC...3'
3'...CAAATTTG(N)6CACG...5'		 blocked
NsbI
FastDigest® fspI (NsbI)		 5'...AAC(N)6GTGCGCA...3'
3'...TTG(N)6CACGCGT...5'		 not blocked
OliI 5'...AACAC(N)4GTGC...3'
3'...TTGTG(N)4CACG...5'		 blocked
RseI
FastDigest® MslI (RseI)		 5'...AACAY(N)4GTGC...3'
3'...TTGTR(N)4CACG...5'		 blocked
Tru1I
FastDigest® Tru1I		 5'...TTAAC(N)6GTGC...3'
3'...AATTG(N)6CACG...5'		 blocked

The recognition sequences of the following Fermentas enzymes may also overlap EcoBI or EcoKI methylated sequence: 

 Home  Search  Contacts  Order  Catalog  Support

catalog@fermentas.com

Updated balandžio 22, 2008 10:19