DpnI: Difference between revisions
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==Purpose== | ==Purpose== | ||
*To digest methylated GATC sites. Useful for removing cell-derived plasmid template from PCR samples. | *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. | ||
*DpnI cleaves '''hemi'''-adenomethylated dam sites 60X more slowly than fully methylated. [http://www.neb.com/nebecomm/products/faqproductR0176.asp#10 NEB FAQ] | |||
*Plasmids derived from Dam+ ''E.coli'' strains will be a substrate | |||
===Relevant protocols=== | ===Relevant protocols=== | ||
Latest revision as of 05:29, 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.
