Common Problems in Magnetic Bead Method Nucleic Acid Extraction

In the process of nucleic acid detection, magnetic bead extraction is widely used because of its high detection sensitivity, automatizable operation, high throughput, safety, and rapidity. However, in the process of nucleic acid extraction, there are more details to pay attention to; an error will lead to the failure of the results; the following to help you organize some common problems and answers. I hope you will be helpful.

Can the efficiency of nucleic acid extraction be improved by increasing the amount of magnetic beads?

When the extraction result is not good, some people may think that increasing the dosage of magnetic beads can absorb more nucleic acids and improve the amount of nucleic acid extraction, but in fact, it isn’t easy to achieve the purpose of this method.

Excessive dosage of magnetic beads will result in their inability to be evenly dispersed into the solution system at each step, reducing their dispersibility and leading to incomplete adsorption, washing, and elution processes. Excessive amounts of magnetic beads will also adsorb more impurities, and the absence of nucleic acid binding sites on the surface of magnetic beads will provide more opportunities for the binding of impurities. In general, the amount of magnetic beads given in the magnetic bead method nucleic acid extraction kit is enough to meet the common application scenarios; if you encounter the problem of low nucleic acid extraction yield, you need to give full consideration to the cleavage and binding, cleaning and elution process.

How to avoid nucleic acid degradation during nucleic acid extraction?

Each step should pay attention to adequate mixing uniformity to avoid the residue of agglomerates affecting the experimental results; in the elution, too long or too high temperature, the nucleic acid may be degraded; samples remember not to repeated freezing and thawing, extracted samples should be careful to save at the right temperature, long-term preservation of as far as possible -80 ℃.

Sample lysis effect is not good, can we increase the amount of lysate used to improve the lysis effect and enhance the extraction effect?

For the magnetic bead method, every increase in the volume of the liquid reduces the chance of collision of the magnetic beads, while reducing the chance of collision of the magnetic beads will lead to a significant decrease in the adsorption rate. So very often, although increasing the lysis solution and washing solution can indeed play a role in enhancing lysis and washing, the core of magnetic bead extraction is the efficiency of magnetic bead adsorption of nucleic acid, can not guarantee the efficiency of magnetic bead collision is not able to guarantee the efficiency of nucleic acid extraction, so simply increase the amount of reagents to improve the extraction effect is not necessarily completely effective.

Nucleic acid extraction purity is not high, there are protein or salt residues, is it better to clean more times?

Nucleic acid extraction purity is not high, can be appropriate to increase the number of times one or two cleaning. If the number of cleaning is too high, it will increase the amount of nucleic acid loss in the cleaning process. In general, the number of cleaning times in 2-4 times is appropriate.

Is it possible to enhance the extraction effect by increasing the amount of sample extracted?

Within the range that the sample has high nucleic acid content and can be fully cleaved by the lysate, increasing the sample volume can increase the extraction effect, on the contrary, if the lysate is insufficient and the sample volume is too much, it may affect the cleavage effect, which may lead to the decrease of the quality of the extraction. However, for samples with low nucleic acid content, increasing the sample volume will instead introduce more impurities and reduce the quality of the final nucleic acid extraction. For such samples, it is recommended to go through the enrichment or concentration step in the pre-treatment before starting the extraction.

Why is the effect of magnetic beads, which was originally very good, reduced after changing a kit?

There are many kinds of magnetic beads, different particle size, different dispersion, different magnetic response time, different encapsulation base matrix, different outer layer modification functional groups, different encapsulation density, different functional group arm lengths, which will lead to a great difference in the characteristics of magnetic beads.

So the experiments and systems adapted to different magnetic beads are also different. The magnetic beads that perform well in the original system are the best combination after a series of screening and method mapping. If the magnetic beads are changed into a new system, it is normal to see a decrease in extraction effect. In most cases, the magnetic beads and reagent systems have to do a certain amount of time with the adjustment.

The extraction effect is not as good as a certain kit; are the magnetic beads bad?

Many users in the process of screening magnetic beads in the already mature reagent system, simply equal amount of replacement of magnetic beads, used to compare the effect of magnetic beads. It is easy to conclude that the effect of a certain bead is not good, but in fact, because different beads are suitable for different systems and dosages, often need to be adjusted to get better extraction results.

Supplier Introduction

Shanghai Lingjun Biotechnology Co., Ltd. was established in 2016 which is a professional manufacturer of biomagnetic materials and nucleic acid extraction reagents.

We have rich experience in nucleic acid extraction and purification, protein purification, cell separation, chemiluminescence and other technical fields.

Our products are widely used in many fields, such as medical testing, genetic testing, university research, genetic breeding, and so on. We not only provide products but also can undertake OEM, ODM, and other needs. If you have related needs, please feel free to contact us at sales01@lingjunbio.com.