Difference between revisions of "DpnI"

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==Purpose==
 
==Purpose==
*To digest methylated GATC sites.  Useful for removing cell-derived plasmid template from PCR samples.
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*To digest (Adeno) methylated GATC sites.  Useful for removing cell-derived plasmid template from PCR samples.
  
 
==Procurement==
 
==Procurement==
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*Incubate at 37°C.
 
*Incubate at 37°C.
 
*Heat inactivate by incubating at 80°C for 20 minutes.
 
*Heat inactivate by incubating at 80°C for 20 minutes.
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*DpnI cleaves '''hemi'''-adenomethylated dam sites 60X more slowly than fully methylated. [http://www.neb.com/nebecomm/products/faqproductR0176.asp#10 NEB FAQ]
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*Plasmids derived from Dam+ ''E.coli'' strains will be a substrate
  
 
===Relevant protocols===
 
===Relevant protocols===

Latest revision as of 04:29, 3 July 2012

Purpose

  • To digest (Adeno) methylated GATC sites. Useful for removing cell-derived plasmid template from PCR samples.

Procurement

  • Can be ordered from NEB.
  • Typically stored at -20°C.

Use

  • Use in NEBuffer 4. However, can be added directly to PCR sample.
  • Incubate at 37°C.
  • Heat inactivate by incubating at 80°C for 20 minutes.
  • DpnI cleaves hemi-adenomethylated dam sites 60X more slowly than fully methylated. NEB FAQ
  • Plasmids derived from Dam+ E.coli strains will be a substrate

Relevant protocols

Safety

  • None known.

References

Contact