Relative Evaluation of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Concentrating On Nucleic Acid Extraction.
(LNJNbio Polystyrene Microspheres)
In the area of modern biotechnology, microsphere materials are widely utilized in the removal and purification of DNA and RNA because of their high certain area, excellent chemical stability and functionalized surface buildings. Among them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are just one of both most extensively examined and used materials. This write-up is given with technological assistance and information analysis by Shanghai Lingjun Biotechnology Co., Ltd., aiming to methodically compare the performance distinctions of these two kinds of products in the procedure of nucleic acid extraction, covering crucial indicators such as their physicochemical properties, surface alteration capability, binding effectiveness and healing rate, and show their applicable situations through speculative data.
Polystyrene microspheres are homogeneous polymer fragments polymerized from styrene monomers with excellent thermal security and mechanical strength. Its surface is a non-polar framework and generally does not have active functional teams. Consequently, when it is straight used for nucleic acid binding, it requires to depend on electrostatic adsorption or hydrophobic activity for molecular fixation. Polystyrene carboxyl microspheres introduce carboxyl useful teams (– COOH) on the basis of PS microspheres, making their surface with the ability of more chemical coupling. These carboxyl groups can be covalently bonded to nucleic acid probes, healthy proteins or other ligands with amino teams via activation systems such as EDC/NHS, therefore achieving more steady molecular addiction. Consequently, from a structural viewpoint, CPS microspheres have a lot more benefits in functionalization potential.
Nucleic acid extraction typically consists of actions such as cell lysis, nucleic acid release, nucleic acid binding to solid phase providers, washing to get rid of pollutants and eluting target nucleic acids. In this system, microspheres play a core role as strong stage service providers. PS microspheres mostly count on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding efficiency has to do with 60 ~ 70%, however the elution performance is low, only 40 ~ 50%. In contrast, CPS microspheres can not only use electrostatic effects however likewise accomplish even more solid fixation with covalent bonding, reducing the loss of nucleic acids during the washing procedure. Its binding efficiency can get to 85 ~ 95%, and the elution performance is also boosted to 70 ~ 80%. Furthermore, CPS microspheres are likewise substantially better than PS microspheres in terms of anti-interference ability and reusability.
In order to validate the performance distinctions between both microspheres in actual procedure, Shanghai Lingjun Biotechnology Co., Ltd. performed RNA removal experiments. The experimental examples were derived from HEK293 cells. After pretreatment with standard Tris-HCl barrier and proteinase K, 5 mg/mL PS and CPS microspheres were utilized for removal. The outcomes revealed that the ordinary RNA yield extracted 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 boosted to 132 ng/ μL, the A260/A280 proportion was close to the perfect value of 1.91, and the RIN value got to 8.1. Although the procedure time of CPS microspheres is slightly longer (28 mins vs. 25 minutes) and the cost is higher (28 yuan vs. 18 yuan/time), its removal high quality is dramatically enhanced, and it is more suitable for high-sensitivity detection, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the viewpoint of application circumstances, PS microspheres are suitable for large-scale screening tasks and initial enrichment with reduced needs for binding uniqueness because of their inexpensive and easy operation. Nevertheless, their nucleic acid binding capacity is weak and quickly impacted by salt ion concentration, making them inappropriate for long-term storage or repeated use. On the other hand, CPS microspheres are suitable for trace sample removal because of their rich surface area useful groups, which help with additional functionalization and can be made use of to create magnetic bead detection packages and automated nucleic acid extraction systems. Although its prep work procedure is relatively intricate and the price is fairly high, it shows stronger adaptability in scientific research and scientific applications with strict requirements on nucleic acid removal efficiency and purity.
With the fast advancement of molecular medical diagnosis, genetics editing, fluid biopsy and other areas, greater demands are placed on the efficiency, pureness and automation of nucleic acid removal. Polystyrene carboxyl microspheres are slowly replacing traditional PS microspheres due to their superb binding performance and functionalizable attributes, becoming the core selection of a new generation of nucleic acid extraction products. Shanghai Lingjun Biotechnology Co., Ltd. is also continuously optimizing the fragment dimension distribution, surface thickness and functionalization efficiency of CPS microspheres and creating matching magnetic composite microsphere products to satisfy the requirements of professional diagnosis, scientific research institutions and commercial clients for premium nucleic acid extraction solutions.
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