Primer Resuspension: A Complete Guide for Molecular Biologists
Our primer resuspension calculator is an essential tool for molecular biologists working with synthetic oligonucleotides. Whether you're preparing primers for PCR, sequencing, or other applications, proper resuspension is critical for experimental success. This guide covers everything from basic oligo resuspension techniques to advanced concentration calculations.
Why Primer Resuspension Matters
Synthetic oligonucleotides typically arrive lyophilized (freeze-dried) and require precise resuspension to achieve accurate concentrations. Our oligo resuspension calculator helps you:
- Convert nmoles to practical working concentrations
- Account for primer length in molecular weight calculations
- Generate dilution schemes for different applications
- Match manufacturer-specific recommendations (IDT, Sigma, Eurofins)
Using the IDT Primer Dilution Calculator
Our IDT primer dilution calculator is specifically calibrated for Integrated DNA Technologies oligos, but works equally well with primers from other suppliers. The calculator accounts for:
| Parameter | Default Value | Adjustable | Notes |
|---|---|---|---|
| Average MW per base | 330 g/mol | Yes | Standard for DNA |
| Extinction coefficient | 0.027 μg/μL per OD260 | No | Standard value |
| Working concentration | 100 μM | Yes | Typical for PCR |
When to Use Different Concentrations
According to research from the National Institutes of Health, primer concentration significantly impacts molecular biology applications:
- Standard PCR: 0.1-0.5 μM final concentration
- qPCR: 0.2-0.9 μM (optimize for each primer pair)
- Sequencing: 0.5-1.0 μM
- Site-directed mutagenesis: 0.1-0.3 μM
Important Considerations
Always verify primer concentrations by spectrophotometry when possible. Our resuspension calculator IDT provides estimates that should be confirmed experimentally for critical applications.