Magni:Protocols/Digestion

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search

Lorenzo Pasotti (Talk | contribs)
(New page: {{Magni}} <div style="padding: 10px; width: 720px; border: 5px solid #000000;"> = Digestion = To perform a double digest with the Roche enzymes, add to a 0.2-ml PCR tube: **1000 ng of DNA...)
Next diff →

Revision as of 17:00, 9 September 2012

Home        Contact        Lab Members        Publications        Research        Internal        Protocols        Data       


Digestion

To perform a double digest with the Roche enzymes, add to a 0.2-ml PCR tube:

    • 1000 ng of DNA or up to 20.5 ul of DNA (if performing the excision of an insert for a ligation, e.g. E-P, X-P, E-S) or
    • 500-700 ng of DNA or up to 20.5 ul of DNA (if opening a plasmid E-X or S-P for a ligation) or
    • 50-1000 ng of DNA or up to 20.5 ul of DNA (if performing a screening)
  • Milli-Q water up to 20.5 ul.
  • 2.5 ul of buffer H
  • 1 ul of the first enzyme
  • 1 ul of the second enzyme
Mix well and incubate in the thermocycler (Digestion program) for 2-4 hours if using the products for a ligation, or for 10-20 minutes if using the products for a screening.
Personal tools