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Resuspension Calculator

Created by Komal Rafay
Reviewed by Purnima Singh, PhD and Rijk de Wet
Last updated: Jan 18, 2024


Suppose we were to tell you that we have a resuspension calculator that will take out all the guessing from dissolving oligos and help you with accurate measurements to prepare your stock solution! How would that make you feel?

If that makes you feel optimistic, then you should also know we have many exciting topics for you to read about — like primer and cell resuspension or what a resuspension buffer is. We will also explain the capabilities of our oligo resuspension calculator, elaborating on the resuspension formula as we go.

What is resuspension?

In a solution, the substances that tend to settle or sediment at the bottom must be mixed or agitated. This process of remixing is called resuspension. In chemistry, this solution can be either gas or liquid. This is typically done to redistribute the particles or molecules of the substance evenly throughout the liquid, allowing for more accurate analysis or use.

You may come across many terms surrounding resuspension, like:

  • Primer resuspension;
  • Oligo resuspension;
  • Cell resuspension; and
  • Resuspension buffer.

We will be talking about them later in the article, so stay tuned!

Oligo resuspension calculator

An oligo resuspension calculator is a tool used to determine the volume of the diluent needed to dissolve a given amount of oligonucleotides (short DNA or RNA molecules) to a desired concentration.

Our calculator is a convenient tool of its kind that takes into consideration the amount of oligos, the desired concentration, and the volume of the diluent. And the most interesting thing about our tool — as long as you have any two of the three mentioned quantities, it will automatically determine the third for you.

So how do you determine the volume of diluent using our resuspension calculator? Follow the steps below, and you will have your answer.

  1. Input the amount of oligonucleotides. The default unit is nmol\text{nmol} (nanomoles), but you have the option to change to other mole based units.
  2. Next, input the desired concentration of the solution you want for resuspension. The default unit is μM\text{μM} (micromolar), which is one millionth of a mole per liter. You may change it to other available concentration units.
  3. The result is the required volume of the diluent needed to prepare the resuspension solution. The default unit is μL\text{μL}, but you can change it to any other volume measuring unit.

Let's take an example, say you have a 60 nmol60 \text{ nmol} oligo, and the desired solution concentration is 40 μM40 \text{ μM} — what would be the diluent volume to resuspend the oligos? In the calculator, you'd enter 6060 as the amount of oligo and 6060 as the desired concentration. The result would be 1, ⁣500 μL1,\!500 \text{ μL} as the required volume of diluent.

💡 We also have a reconstitution calculator that helps prepare a solution from a powder by determining the reconstitution concentration of a solution.

Cell resuspension solution

A cell resuspension solution is a liquid that resuspends cells that are clumped together after being harvested from a culture. This solution generally comprises a buffer to regulate pH and components such as salts, sugars, and proteins to provide nutrients.

You may use it to resuspend cells, proteins, DNA, and other biological materials. The cell resuspension solution helps improve the precision of experiments that demand homogeneous solutions.

Resuspension formula

The composition of the resuspension solution may differ based on what type of cell you need to resuspend. So, it is time to check out the resuspension formula:

Required volume of diluent [μL]=Amount of oligo [nmol]×1000Desired concentration [μM]\footnotesize \text {Required volume of diluent [μL]} \\[0.5em] = \frac{\text {Amount of oligo [nmol]} \times 1000}{\text {Desired concentration [μM]}}

This means all you have to do is take the amount of oligo in nanomol, multiply it by 1000, and divide it by the desired concentration of your resuspension solution in micromolar, to estimate the solvent volume in microliter.

For example, if you want to resuspend 10 nmol10 \text{ nmol} of oligo in TE buffer to a concentration of 100 μM100 \text{ μM}, you need to determine the amount of the buffer. Let's substitute the values in the resuspension formula:

Required volume of diluent=10×1,000/100=100 μL\footnotesize \text {Required volume of diluent} \\ = 10 \times 1,\!000 / 100 \\ = 100 \text{ μL}

To ensure complete resuspension, pipet or vortex the buffer after adding the oligo.

💡 Might we suggest trying out our alligation calculator? It can calculate the proportions of two solutions to be mixed to obtain a desired third solution!

Significance of oligo resuspension

A uniform distribution of oligonucleotides in a solution, achieved by resuspension, is a prerequisite for many molecular biology applications like:

  • Polymerase chain reaction (PCR): A lab technique to amplify segments of DNA;
  • Sequencing and DNA sequencing: The process of determining the precise order of nucleotides; and
  • Gene expression analysis: The process of studying the genetic information composed in a cell.

Oligonucleotides are highly sensitive to environmental factors, making resuspension a difficult process. Hence the need to dissolve them in a buffer solution to ensure an optimal environment.

💡 Check out the annealing temperature calculator, where you will learn all about PCR, annealing temperature, PCR thermal cycle, and much more.

What is resuspension buffer?

A resuspension buffer is a solution we use to resuspend and dissolve biological samples, such as cells or DNA. The buffer's composition comprises salts, detergents, and other stabilizing agents that preserve the sample's integrity and prevent degradation. The design of the resuspension buffer may vary depending on the specific application.

FAQ

Is primer resuspension different from oligo resuspension?

Oligo and primer resuspension are different from each other, but only in terms of their applications and the types of genetic sequences to be resuspended. Primer resuspension is the process of resuspending primers in a buffer solution. Depending on usage, the buffer solution is either TE buffer or nuclease-free water. Whereas oligo resuspension involves dissolving or resuspending oligonucleotides in a buffer solution. Here, the buffer solution can also be TE or nuclease-free water.

What does a cell resuspension solution do?

A cell resuspension solution rehydrates and resuspends cells that have been clumped or frozen. It also helps in breaking up cell clots, maintaining cell viability, and preventing cell damage during the resuspension process. A resuspension solution generally contains salts, buffers, and nutrients that help to maintain the proper pH and osmotic balance for the cells.

How much diluent do I need to make a 1600 micromolar solution?

You will need 62.5 μL (microliters) of diluent to prepare a resuspension solution with 100 nmol of oligos and a concentration of 1600 μM.

You can easily determine the diluent volume by following these simple steps:

  1. Multiply the amount of oligo by 1000.
  2. Divide the result by the desired concentration of the solution.
  3. The result is the volume of the diluent.
Komal Rafay
Oligo amount
nmol
Desired concentration
μM
Diluent volume required
µl
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