Methylation-sensitive restriction enzymes (MSREs)

Methylation-Sensitive Restriction Enzymes (MSREs) play a role in the analysis of methylated DNA, as they are used to analyze the methylation status of cytosine residues in CpG sequences. These restriction enzymes, as their name implies, are not able to cleave methylated-cytosine residues, leaving methylated DNA intact. MSREs cleave DNA at specific unmethylated-cytosine residues and DNA is then amplified by PCR following digestion. Because unmethylated DNA is digested by MSREs, the only amplification products detected are methylated DNA. It is important to fully digest the DNA before PCR amplification to generate accurate data.

The table below lists MSREs that may be useful for mammalian epigenetics analyses when used with CpG methylase.

Restriction EnzymeRecognition Sequence
Aat II GACGT↓C
Acc II CG↓CG
Aor13H I T↓CCGGA
Aor51H I AGC↓GCT
BspT104 I TT↓CGAA
BssH II G↓CGCGC
Cfr10 I R↓CCGGY
Cla I AT↓CGAT
Cpo I CG↓GWCCG
Eco52 I C↓GGCCG
Hae II RGCGC↓Y
Hap II C↓CGG
Hha I GCG↓C
Mlu I A↓CGCGT
Nae I GCC↓GGC
Not I GC↓GGCCGC
Nru I TCG↓CGA
Nsb I TGC↓GCA
PmaC I CAC↓GTG
Psp1406 I AA↓CGTT
Pvu I CGAT↓CG
Sac II CCGC↓GG
Sal I G↓TCGAC
Sma I CCC↓GGG
SnaB I TAC↓GTA