Lyophilised Oligo Reconstitution Calculator
Enter the amount of lyophilised oligonucleotide from your spec sheet and the concentration you need. The calculator tells you exactly how much TE buffer or nuclease-free water to add.
Oligo Details
Check your oligo spec sheet for the yield in µmol (common synthesis scales: 0.01, 0.025, 0.05, 0.2, 1.0 µmol)
Estimated MW: ~6,600 Da (using ~330 Da per nucleotide average)
100 µM is a common stock concentration for primers
Reconstitution Recipe
| Parameter | Value |
|---|---|
| Oligo amount | 0.0250 µmol (0.1650 mg) |
| Molecular weight | 6,600 Da(estimated) |
| Target concentration | 100 µM |
| Volume to add | 250.0 µL |
About Oligo Reconstitution
Synthetic oligonucleotides are typically supplied as a lyophilised (freeze-dried) pellet. Before use, they must be reconstituted in an appropriate solvent to create a stock solution at a known concentration.
How the Calculation Works
The reconstitution volume is calculated from the simple relationship:
When the amount is given in mass (mg), it is first converted to µmol using the molecular weight: µmol = mass (mg) × 1000 ÷ MW (Da). If you don’t know the exact MW, an estimate of ~330 Da per nucleotide is used.
Choosing a Solvent
TE buffer (10 mM Tris-HCl, 0.1 mM EDTA, pH 8.0) is recommended for long-term storage. The slightly alkaline pH prevents depurination, and EDTA chelates metal ions that can catalyse degradation. Nuclease-free water is a good alternative when EDTA might interfere with downstream applications (e.g. certain PCR mixes or enzymatic reactions).
Oligos in Microfluidics
Oligonucleotides are widely used in microfluidic platforms for on-chip PCR, isothermal amplification (LAMP, RPA), hybridisation assays, and aptamer-based biosensors. Precise reconstitution is critical — even small concentration errors can affect assay sensitivity and reproducibility in low-volume microfluidic channels.
Storage Recommendations
Store reconstituted oligos at −20 °C in single-use aliquots. Avoid repeated freeze-thaw cycles. Lyophilised oligos (unopened) are stable at room temperature for several months and at −20 °C for years.
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