Resuspension Calculator
Calculate volumes and concentrations for oligonucleotide resuspension in molecular biology applications
Oligonucleotide Resuspension
Amount of oligonucleotide to resuspend
Target concentration for final solution
Resuspension Results
Resuspension Formula: Volume (μL) = (Amount (nmol) × 1000) / Concentration (μM)
Buffer Recommendation: Unknown
Reason: Enter concentration
Application Recommendations
Example Calculation
PCR Primer Resuspension
Application: Forward primer for gene amplification
Oligo amount: 60 nmol (from synthesis)
Desired stock concentration: 100 μM
Intended use: PCR reactions (working concentration 10 μM)
Step-by-Step Calculation
1. Apply resuspension formula: Volume = (Amount × 1000) / Concentration
2. Substitute values: Volume = (60 nmol × 1000) / 100 μM
3. Calculate: Volume = 60,000 / 100 = 600 μL
Result: Add 600 μL of TE buffer to create 100 μM stock solution
Usage: Dilute 1:10 for 10 μM working solution in PCR
Resuspension Process
Calculate Volume
Use formula to determine buffer volume
Volume = (Amount × 1000) / Concentration
Add Buffer
Add calculated volume of buffer/water
Use TE buffer or nuclease-free water
Mix Thoroughly
Vortex or pipette to ensure dissolution
Avoid introducing air bubbles
Common Applications
PCR primers: 10-100 μM stock concentrations
qPCR probes: 1-10 μM working concentrations
Sequencing primers: 3.2 μM standard concentration
Cloning vectors: Variable based on application
Understanding Oligonucleotide Resuspension
What is Resuspension?
Resuspension is the process of dissolving lyophilized (freeze-dried) oligonucleotides in an appropriate buffer or solvent to create a stock solution of known concentration. This is essential for accurate dosing in molecular biology applications.
Why is Proper Resuspension Important?
- •Ensures accurate concentration for experimental reproducibility
- •Prevents degradation through proper pH and ionic conditions
- •Allows for precise dilutions and aliquoting
- •Facilitates long-term storage and stability
Resuspension Formula
Volume (μL) = (Amount (nmol) × 1000) / Concentration (μM)
- Volume: Required buffer volume in microliters
- Amount: Oligonucleotide quantity in nanomoles
- 1000: Conversion factor (nmol to μmol)
- Concentration: Desired final concentration in micromolar
Tip: Always centrifuge the oligo tube before opening to ensure all material is at the bottom
Buffer Selection Guidelines
TE Buffer
Composition: 10 mM Tris-HCl, 1 mM EDTA, pH 8.0
Use for: Long-term storage, standard applications, protection against nucleases
Nuclease-Free Water
Composition: Sterile, DNase/RNase-free water
Use for: High concentration stocks, applications sensitive to buffer components
Custom Buffers
Examples: Low-salt TE, Tris buffer without EDTA
Use for: Specific applications requiring particular ionic conditions