Molecular Research Center develops Formazol for RNA solubilization and protection

FORMAzol®
Cat. No. FO 121
Store at 4 C. Use within two years from date of purchase.

PRODUCT DESCRIPTION
FORMAzol is a specially purified and stabilized formamide for use as an RNA solubilizer. Unlike any other commercially available formamide, FORMAzol is stable and can be used (without additional purification) for RNA solubilization for two years when stored at 4 C or -20 C. Stabilized formamide has several advantages over water as an RNA solubilization agent:

RNA solubilized in FORMAzol is protected from degradation by RNase.

RNA solubilized in FORMAzol can be stored for at least two years at -20 C instead of at -70 C as required for RNA solubilized in water.

A high sample volume (up to 50% of a slot volume) can be applied to a formaldehyde-agarose gel.
[P.Chomczynski (1992), Nucleic Acid Res 20, 3791-3792].

APPLICATION NOTES

RNA SOLUBILIZATION
Drying of RNA to evaporate residual amount of ethanol is not recommended. Add 100 µl of FORMAzol to a 40 - 400 µg RNA sample. Solubilize RNA by repetitive pipetting and incubation at 50 - 55 C for 10 - 15 minutes.

RNA PRECIPITATION
FORMAzol inhibits reverse transcriptase. For use in RT-PCR, precipitate RNA from FORMAzol by the addition of 4 volumes of ethanol. Store samples 3-5 minutes at room temperature and centrifuge at 10,000 g for 5 minutes. When samples contain < 20 µg RNA, add NaCl (do not use sodium acetate) to final concentration of 0.2 M followed by 4 volumes of ethanol.

RNA QUANTITATION
Mix an aliquot of the RNA sample (3 - 10 µl) with 1.0 ml water or 1-3 mM Na₂HPO₄ and measure its A₂₆₀ against a blank sample containing 1.0 ml water and the same volume of FORMAzol as the RNA sample.

FORMALDEHYDE-GEL ELECTROPHORESIS
Prepare a formaldehyde reaction solution containing: water (87 µl), formaldehyde (81 µl), bromophenol blue (0.25 mg/ml) in 50% glycerol (48 µl) and 20X MOPS buffer (24 µl). The formaldehyde reaction solution is highly unstable and should be prepared immediately before use. Mix equal volumes of the RNA sample solubilized in FORMAzol and the formaldehyde reaction solution and incubate at 55 C for 15 minutes. Apply this mixture on a formaldehyde-agarose gel and perform the electrophoresis according to your standard protocol.