Comparative Evaluation of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Focusing on Nucleic Acid Extraction.
(LNJNbio Polystyrene Microspheres)
In the field of modern biotechnology, microsphere products are widely utilized in the removal and filtration of DNA and RNA due to their high certain area, good chemical security and functionalized surface residential or commercial properties. Among them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are one of the two most commonly examined and applied products. This write-up is offered with technological support and information analysis by Shanghai Lingjun Biotechnology Co., Ltd., intending to methodically compare the efficiency differences of these 2 kinds of materials in the procedure of nucleic acid extraction, covering essential indications such as their physicochemical buildings, surface area adjustment ability, binding efficiency and recuperation price, and illustrate their applicable situations with experimental information.
Polystyrene microspheres are homogeneous polymer bits polymerized from styrene monomers with great thermal stability and mechanical stamina. Its surface is a non-polar framework and typically does not have energetic functional groups. As a result, when it is straight utilized for nucleic acid binding, it needs to depend on electrostatic adsorption or hydrophobic activity for molecular fixation. Polystyrene carboxyl microspheres introduce carboxyl functional groups (– COOH) on the basis of PS microspheres, making their surface efficient in more chemical coupling. These carboxyl groups can be covalently bonded to nucleic acid probes, proteins or other ligands with amino teams with activation systems such as EDC/NHS, thereby achieving extra steady molecular addiction. As a result, from a structural viewpoint, CPS microspheres have more advantages in functionalization potential.
Nucleic acid extraction generally includes steps such as cell lysis, nucleic acid release, nucleic acid binding to strong stage service providers, washing to get rid of contaminations and eluting target nucleic acids. In this system, microspheres play a core duty as strong phase carriers. PS microspheres mainly rely on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding efficiency is about 60 ~ 70%, but the elution performance is reduced, only 40 ~ 50%. On the other hand, CPS microspheres can not just make use of electrostatic impacts but additionally accomplish even more solid addiction via covalent bonding, minimizing the loss of nucleic acids during the washing process. Its binding performance can reach 85 ~ 95%, and the elution performance is also increased to 70 ~ 80%. Furthermore, CPS microspheres are also considerably far better than PS microspheres in regards to anti-interference capability and reusability.
In order to verify the performance differences between the two microspheres in real operation, Shanghai Lingjun Biotechnology Co., Ltd. conducted RNA extraction experiments. The speculative samples were originated from HEK293 cells. After pretreatment with conventional Tris-HCl barrier and proteinase K, 5 mg/mL PS and CPS microspheres were used for removal. The outcomes showed that the average RNA yield drawn out by PS microspheres was 85 ng/ μL, the A260/A280 ratio was 1.82, and the RIN value was 7.2, while the RNA return of CPS microspheres was increased to 132 ng/ μL, the A260/A280 ratio was close to the perfect value of 1.91, and the RIN worth reached 8.1. Although the operation time of CPS microspheres is somewhat longer (28 minutes vs. 25 minutes) and the cost is higher (28 yuan vs. 18 yuan/time), its removal top quality is considerably improved, and it is preferable for high-sensitivity detection, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the viewpoint of application situations, PS microspheres are suitable for massive screening projects and preliminary enrichment with reduced needs for binding uniqueness as a result of their affordable and basic procedure. Nonetheless, their nucleic acid binding capability is weak and quickly influenced by salt ion concentration, making them unsuitable for long-term storage or repeated usage. On the other hand, CPS microspheres appropriate for trace sample removal due to their abundant surface practical groups, which help with more functionalization and can be utilized to construct magnetic bead discovery sets and automated nucleic acid removal platforms. Although its preparation procedure is relatively intricate and the cost is fairly high, it reveals stronger adaptability in clinical study and scientific applications with strict demands on nucleic acid removal efficiency and pureness.
With the quick growth of molecular diagnosis, genetics editing and enhancing, fluid biopsy and other fields, higher requirements are put on the performance, purity and automation of nucleic acid extraction. Polystyrene carboxyl microspheres are gradually replacing standard PS microspheres because of their exceptional binding performance and functionalizable attributes, coming to be the core option of a new generation of nucleic acid removal products. Shanghai Lingjun Biotechnology Co., Ltd. is also continually enhancing the particle size distribution, surface area thickness and functionalization efficiency of CPS microspheres and creating matching magnetic composite microsphere items to fulfill the requirements of professional diagnosis, clinical research organizations and industrial consumers for top quality nucleic acid extraction remedies.
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