Formamide
Formamide (HCONH2, CAS 75-12-7) is the simplest amide of formic acid. It is a colourless, hygroscopic, water-miscible liquid used mainly as a polar solvent and as a nucleic acid denaturant in molecular biology. It lowers the melting temperature of DNA and RNA duplexes, so it is common in hybridisation, sequencing and electrophoresis buffers. It is classified as a reproductive toxicant and requires careful handling.
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Formamide
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Formamide (IUPAC name: methanamide) has the formula HCONH2 and a molar mass of about 45.04 g/mol. It is a colourless, nearly odourless, hygroscopic liquid that is fully miscible with water. Its high polarity and high dielectric constant make it an effective solvent for many salts and polar compounds.
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What is formamide used for in the laboratory?
In molecular biology, formamide disrupts hydrogen bonding between base pairs. This lowers the melting temperature (Tm) of nucleic acid duplexes. It is therefore used in:
- Northern, Southern and in situ hybridisation buffers, where it allows stringent conditions at lower temperatures.
- Denaturing loading buffers for RNA and sequencing gels.
- Sample preparation for capillary electrophoresis in DNA sequencing and fragment analysis.
It is also used as an industrial solvent and as a chemical intermediate.
Which grade should I specify when purchasing?
For hybridisation and sequencing work, buyers normally specify molecular biology or deionised grade, tested for nuclease activity and low conductivity. Formamide slowly hydrolyses to ammonium formate, and this ionic contamination can affect electrophoresis and hybridisation results. Suppliers often recommend storing it cold and in aliquots. Check the CoA for purity (often assayed by GC), water content and conductivity. Reagent grades such as ACS are suitable for general solvent use.
What are the safety and regulatory considerations?
Under GHS/CLP, formamide is classified as a reproductive toxicant, Category 1B (H360D: may damage the unborn child). In the EU it is on the REACH Candidate List of Substances of Very High Concern. It should be handled in a fume hood with suitable gloves and eye protection, following the supplier's SDS. Procurement and EHS teams should check the current national and regional restrictions that apply to their use.
Related concepts
Formamide is often discussed alongside other denaturants such as urea and dimethyl sulfoxide (DMSO). It is used with hybridisation buffers such as SSC and with nucleic acid melting temperature (Tm) calculations. Related amide solvents include N,N-dimethylformamide (DMF).
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