Jacobs:Protocol RNA Sample Loading Buffer

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Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1989), p. 7.43-7.45, 7.51
==Contact==
==Contact==

Revision as of 22:24, 16 June 2008

Contents

Overview

RNA sample loading buffer is formulated for use in RNA sample preparation for denaturing formaldehydeagarose gel electrophoresis. The solution contains tracking dyes and ethidium bromide, so gel loading buffer and ethidium bromide staining are not required.

Materials

  • Composition:
    • 62.5 % deionized formamide
    • 1.14 M formaldehyde
    • 1.25X MOPS-EDTA-sodium acetate buffer (Product No. M 5755, diluted 1:8)
    • 200 μg/ml bromphenol blue
    • 200 μg/ml xylene cyanole
    • 50 μg/ml ethidium bromide (See note below)
  • Suitable for use in formaldehyde-agarose gel electrophoresis of RNA.
    • RNase: None detected
    • Store at 0 to −20 °C


Note: This solution contains ethidium bromide that can interfere with transfer of RNA to nylon or nitrocellulose membranes.

Procedure

Add loading buffer to sample in ratio of 1:2 to 1:5. Just before loading, heat to 65 °C for 10 minutes, then chill on ice.

References

Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1989), p. 7.43-7.45, 7.51

Contact

  • Originally prepared by CRJ-EJC 1/3/06


or instead, discuss this protocol.

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