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How does the adsorption of nucleic acids on the surface of Centrifuge tubes affect downstream applications such as PCR, qPCR or sequencing?

Feb 01,2024

The adsorption of nucleic acids on the surface of Centrifuge tubes can have a significant impact on downstream applications such as PCR, qPCR, or sequencing. Nucleic acid adsorption refers to the binding of DNA or RNA molecules to the surface of the tubes, which can result in the loss or alteration of the nucleic acid samples.
Loss of nucleic acids: When nucleic acids adsorb to the tube surface, they can be lost during various steps of the experimental process. This loss can lead to lower yields of DNA or RNA, making it challenging to obtain sufficient amounts of nucleic acids for downstream applications.
Inhibition of enzymatic reactions: Adsorbed nucleic acids can inhibit enzymatic reactions, such as PCR or qPCR. The adsorbed molecules can act as inhibitors by binding to the polymerases or other critical components involved in the amplification or detection process. This inhibition can result in reduced amplification efficiency, lower sensitivity, or even complete failure in the amplification or detection of the target sequences.
Contaminating effects: Adsorbed nucleic acids can also act as contaminants in downstream applications. These adsorbed molecules can introduce unwanted sequences or impurities that can interfere with the specificity and accuracy of the assays. Contaminating nucleic acids can lead to false-positive or false-negative results, compromising the reliability of the experimental data.
Alteration of nucleic acid properties: The adsorption of nucleic acids on Centrifuge tubes can also cause physical or chemical alterations to the molecules. For example, the adsorbed molecules might undergo shearing or fragmentation, resulting in shorter or modified nucleic acid fragments. These alterations can impact the accuracy and sequencing depth of DNA or RNA sequencing, leading to incomplete or biased sequence information.
To minimize the negative effects of nucleic acid adsorption, certain measures can be taken. Firstly, using nucleic acid-free centrifuge tubes or pre-treatment of the tube surfaces with suitable coatings can reduce adsorption. Additionally, optimizing the sample preparation protocols, including buffering conditions, pH, and the use of carrier molecules like bovine serum albumin (BSA), can help mitigate the loss or inhibition caused by adsorption. Finally, using molecular biology-grade disposable plastics and careful handling of samples can also minimize the risk of nucleic acid adsorption and potential downstream application issues.