Comparative Evaluation of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Concentrating On Nucleic Acid Removal.
(LNJNbio Polystyrene Microspheres)
In the field of contemporary biotechnology, microsphere materials are widely used in the extraction and filtration of DNA and RNA due to their high certain area, great chemical stability and functionalized surface area homes. Amongst them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are among both most commonly examined and applied materials. This article is provided with technical assistance and data evaluation by Shanghai Lingjun Biotechnology Co., Ltd., aiming to methodically contrast the performance differences of these two types of products in the procedure of nucleic acid removal, covering key indications such as their physicochemical properties, surface area modification capacity, binding performance and recuperation rate, and illustrate their relevant scenarios through experimental data.
Polystyrene microspheres are homogeneous polymer fragments polymerized from styrene monomers with great thermal stability and mechanical toughness. Its surface is a non-polar framework and usually does not have active practical teams. As a result, when it is straight used for nucleic acid binding, it requires to count on electrostatic adsorption or hydrophobic activity for molecular fixation. Polystyrene carboxyl microspheres present carboxyl practical groups (– COOH) on the basis of PS microspheres, making their surface area capable of further chemical combining. These carboxyl teams can be covalently bound to nucleic acid probes, proteins or various other ligands with amino groups with activation systems such as EDC/NHS, therefore accomplishing more steady molecular addiction. As a result, from an architectural point of view, CPS microspheres have extra benefits in functionalization possibility.
Nucleic acid extraction typically includes steps such as cell lysis, nucleic acid release, nucleic acid binding to strong phase providers, cleaning to get rid of pollutants and eluting target nucleic acids. In this system, microspheres play a core function as strong phase providers. PS microspheres mostly rely upon electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding efficiency is about 60 ~ 70%, but the elution efficiency is reduced, just 40 ~ 50%. On the other hand, CPS microspheres can not only use electrostatic effects yet additionally accomplish even more strong addiction with covalent bonding, reducing the loss of nucleic acids throughout the cleaning process. Its binding performance can get to 85 ~ 95%, and the elution performance is likewise raised to 70 ~ 80%. In addition, CPS microspheres are additionally significantly much better than PS microspheres in regards to anti-interference capacity and reusability.
In order to verify the efficiency distinctions between both microspheres in real procedure, Shanghai Lingjun Biotechnology Co., Ltd. performed RNA removal experiments. The speculative samples were stemmed from HEK293 cells. After pretreatment with common Tris-HCl barrier and proteinase K, 5 mg/mL PS and CPS microspheres were utilized for extraction. The results revealed that the ordinary RNA yield extracted by PS microspheres was 85 ng/ μL, the A260/A280 ratio was 1.82, and the RIN worth was 7.2, while the RNA yield of CPS microspheres was enhanced to 132 ng/ μL, the A260/A280 proportion was close to the suitable value of 1.91, and the RIN value got to 8.1. Although the operation time of CPS microspheres is somewhat longer (28 mins vs. 25 mins) and the cost is greater (28 yuan vs. 18 yuan/time), its extraction quality is dramatically improved, and it is better for high-sensitivity detection, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the point of view of application situations, PS microspheres appropriate for large-scale screening tasks and initial enrichment with reduced needs for binding uniqueness due to their low cost and basic procedure. Nevertheless, their nucleic acid binding capacity is weak and conveniently impacted by salt ion focus, making them inappropriate for long-lasting storage or duplicated usage. In contrast, CPS microspheres are suitable for trace example removal because of their rich surface area practical groups, which assist in further functionalization and can be utilized to build magnetic grain detection kits and automated nucleic acid extraction systems. Although its preparation process is relatively complex and the cost is reasonably high, it shows more powerful flexibility in clinical research study and professional applications with stringent requirements on nucleic acid extraction efficiency and purity.
With the fast development of molecular medical diagnosis, gene modifying, liquid biopsy and various other fields, greater demands are positioned on the efficiency, purity and automation of nucleic acid removal. Polystyrene carboxyl microspheres are slowly changing typical PS microspheres because of their excellent binding efficiency and functionalizable qualities, becoming the core choice of a brand-new generation of nucleic acid extraction materials. Shanghai Lingjun Biotechnology Co., Ltd. is also constantly enhancing the particle size distribution, surface area density and functionalization efficiency of CPS microspheres and developing matching magnetic composite microsphere items to satisfy the needs of professional medical diagnosis, clinical research institutions and industrial customers for top notch nucleic acid extraction services.
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