Berglund:Large Scale: 10ml

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Overview

Protocol from Mary, as yet untested in our lab.

Materials

  • 20µg/mL linearized plasmid DNA
  • 40mM Tris pH 8.0
  • 10mM DTT
  • 10mM MgCl2
  • 1mM each rNTP
  • 2mM spermidine
  • radioactive nucleotide?
  • T7 RNA poly

Procedure

  1. Incubate at 37°C for 5 hours.
  2. PAGE purify.

Notes

UNTESTED. Consult with Amy before proceeding.

References

  1. Silverman SK and Cech TR. RNA tertiary folding monitored by fluorescence of covalently attached pyrene. Biochemistry. 1999 Oct 26;38(43):14224-37. PubMed ID:10571996 | HubMed [Silverman-Biochemistry-1999]