Berglund:Large Scale: 10ml

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. DOI:10.1021/bi991333f |