10X BPTE electrophoresis buffer

Purpose

Used as the electrophoresis buffer during denaturing (via glyoxal) agarose gel electrophoresis of RNA.

Components

• 100 mM PIPES
• 300 mM Bis-Tris
• 10 mM EDTA

The final pH of this 10x buffer is ~6.5.

Procedure

1. Prepare by adding the following to 90 ml of distilled H₂O
   • 3 g of PIPES (free acid)
   • 6 g of Bis-Tris (free base)
   • 2 ml of 0.5 M EDTA
2. Treat the solution with final concentration of 0.1% DEPC for 1 hour at 37°C
3. Autoclave.

References

• Molecular Cloning