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Products >  Molecular_Biology >  Restriction_Enzymes >  Restriction_Enzymes_c_d >  DpnI

Methylation Dependent Restriction Enzyme - Dpn I

DNA restriction enzymes from Takara Bio are unsurpassed in quality and purity. We’ve been producing them for over 30 years and were the first manufacturers to offer commercially available restriction enzymes in Japan. Each lot of every DNA restriction enzyme undergoes stringent quality control tests. Takara Bio DNA restriction enzymes are supplied with optimized buffers that provide maximum activity during restriction enzyme digestion. Visit our Restriction Enzyme Applications pages for information on how to digest DNA, relative activity in each Universal Buffer, star activity, buffers for double digestion, effects of DNA methylation, how to inactivate enzymes, and more.

Adenine Methylation Analysis with DpnI

Dpn I is a Type IIM restriction enzyme that specifically cleaves DNA containing methylated adenine (mA) in the recognition sequence GmA | TC, also referred to as the dam sequence since it is recognized by Dam methylase. Dpn I does not cleave unmethylated DNA, and will cleave hemimethylated DNA (with only one adenine methylated) 60-fold more slowly than fully adenomethylated recognition sequences. While DpnI shares the same recognition sequence as DpnII, Sau3AI, and MboI, the isoschizomers have different methylation sensitivities. Sau3AI is insensitive to adenomethylation but will not cleave if one or more cytosines in the recognition sequence are methylated. DpnII and MboI do not cleave adenomethylated sequences.
At-A-Glance Documents

Features

GmA|TC 
CT|mAG 
  • Source: E. coli carrying a plasmid encoding the DpnI gene
  • Supplied buffer: T
  • Reaction temperature: 37°C
  • Methylation sensitivity: requires methylation of adenine in GATC recognition sequence. Hemiadenomethylation results in greatly reduced activity. Cleaves DNA prepared from E. coli dam+ strain. Does not cleave PCR-amplified DNA.
  • Substrate for unit definition: pBR322 DNA prepared from dam+ strain
  • Inactivation: fully inactivated by incubation at 70°C for 15 min. Alternatively, add 1/10 volume of 10X Loading Buffer (supplied) and load directly on an agarose gel for analysis.

Composition of 10X Loading Buffer

0.9% SDS
50% glycerol
0.05% Bromophenyl Blue

Store 10X Loading Buffer at room temperature. SDS may precipitate during storage. If precipitates are present, dissolve by incubating briefly at 37° before use.

Quality control

Takara follows a stringent QC process when manufacturing their restriction enzymes. Each lot of every enzyme undergoes four quality tests including: overdigestion, Genome DNA analysis, Ligation-Recutting and pKF3 Cloning.


 
 
Products
New Cat. # Product Package Size Price # of Units Select
1235A Dpn I 1,000 Units $36.00
1235B Dpn I 5,000 Units $153.00
 

 


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