Comparative Analysis of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Focusing on Nucleic Acid Removal.
(LNJNbio Polystyrene Microspheres)
In the field of modern biotechnology, microsphere materials are widely made use of in the extraction and purification of DNA and RNA due to their high certain surface area, good chemical security and functionalized surface area homes. Amongst them, polystyrene (PS) microspheres and their acquired polystyrene carboxyl (CPS) microspheres are one of the two most widely studied and applied products. This article is given with technical support and information evaluation by Shanghai Lingjun Biotechnology Co., Ltd., aiming to systematically compare the efficiency distinctions of these two types of products in the procedure of nucleic acid removal, covering key indicators such as their physicochemical properties, surface area modification capability, binding efficiency and recovery price, and highlight their appropriate circumstances with speculative data.
Polystyrene microspheres are homogeneous polymer particles polymerized from styrene monomers with good thermal stability and mechanical strength. Its surface area is a non-polar framework and usually does not have active functional teams. Consequently, when it is straight used for nucleic acid binding, it requires to rely on electrostatic adsorption or hydrophobic action for molecular fixation. Polystyrene carboxyl microspheres introduce carboxyl functional teams (– COOH) on the basis of PS microspheres, making their surface with the ability of more chemical combining. These carboxyl groups can be covalently bonded to nucleic acid probes, healthy proteins or various other ligands with amino groups with activation systems such as EDC/NHS, consequently accomplishing more stable molecular addiction. Consequently, from an architectural perspective, CPS microspheres have much more benefits in functionalization possibility.
Nucleic acid removal typically includes steps such as cell lysis, nucleic acid launch, nucleic acid binding to strong phase service providers, washing to get rid of impurities and eluting target nucleic acids. In this system, microspheres play a core role as solid phase providers. PS microspheres generally rely on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding effectiveness has to do with 60 ~ 70%, however the elution effectiveness is reduced, only 40 ~ 50%. On the other hand, CPS microspheres can not just make use of electrostatic effects yet also accomplish even more strong fixation with covalent bonding, decreasing the loss of nucleic acids during the washing procedure. Its binding effectiveness can reach 85 ~ 95%, and the elution effectiveness is also boosted to 70 ~ 80%. In addition, CPS microspheres are likewise dramatically far better than PS microspheres in terms of anti-interference capability and reusability.
In order to verify the efficiency distinctions in between the two microspheres in real operation, Shanghai Lingjun Biotechnology Co., Ltd. performed RNA removal experiments. The speculative examples were derived from HEK293 cells. After pretreatment with typical Tris-HCl barrier and proteinase K, 5 mg/mL PS and CPS microspheres were made use of for extraction. The results showed that the ordinary RNA return removed by PS microspheres was 85 ng/ μL, the A260/A280 ratio was 1.82, and the RIN value was 7.2, while the RNA yield of CPS microspheres was boosted to 132 ng/ μL, the A260/A280 proportion was close to the excellent value of 1.91, and the RIN worth got to 8.1. Although the procedure time of CPS microspheres is a little longer (28 minutes vs. 25 minutes) and the price is greater (28 yuan vs. 18 yuan/time), its extraction high quality is dramatically improved, and it is more suitable for high-sensitivity detection, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the point of view of application scenarios, PS microspheres are suitable for large-scale screening projects and preliminary enrichment with reduced demands for binding specificity as a result of their low cost and straightforward operation. Nevertheless, their nucleic acid binding ability is weak and quickly affected by salt ion concentration, making them inappropriate for long-term storage or repeated usage. On the other hand, CPS microspheres appropriate for trace example removal due to their rich surface practical teams, which assist in more functionalization and can be utilized to build magnetic grain detection packages and automated nucleic acid removal platforms. Although its preparation process is reasonably complicated and the cost is reasonably high, it reveals more powerful flexibility in clinical research study and scientific applications with rigorous requirements on nucleic acid removal effectiveness and purity.
With the fast development of molecular medical diagnosis, gene editing and enhancing, fluid biopsy and various other areas, greater needs are positioned on the performance, pureness and automation of nucleic acid extraction. Polystyrene carboxyl microspheres are gradually replacing traditional PS microspheres as a result of their exceptional binding performance and functionalizable characteristics, becoming the core option of a brand-new generation of nucleic acid extraction products. Shanghai Lingjun Biotechnology Co., Ltd. is likewise constantly enhancing the particle dimension circulation, surface area thickness and functionalization performance of CPS microspheres and creating matching magnetic composite microsphere items to meet the requirements of professional medical diagnosis, clinical study organizations and commercial clients for top quality nucleic acid extraction services.
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