Relative Analysis of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Focusing on Nucleic Acid Extraction.
(LNJNbio Polystyrene Microspheres)
In the area of modern biotechnology, microsphere materials are commonly used in the removal and purification of DNA and RNA due to their high specific surface area, great chemical stability and functionalized surface buildings. Among them, polystyrene (PS) microspheres and their acquired polystyrene carboxyl (CPS) microspheres are one of the two most widely studied and used materials. This short article is given with technological support and information evaluation by Shanghai Lingjun Biotechnology Co., Ltd., aiming to systematically contrast the performance differences of these 2 types of products in the procedure of nucleic acid extraction, covering essential signs such as their physicochemical properties, surface area modification capacity, binding efficiency and healing price, and highlight their appropriate circumstances with experimental data.
Polystyrene microspheres are uniform polymer particles polymerized from styrene monomers with excellent thermal stability and mechanical toughness. Its surface is a non-polar structure and generally does not have energetic practical teams. For that reason, when it is directly utilized for nucleic acid binding, it needs to rely on electrostatic adsorption or hydrophobic activity for molecular addiction. Polystyrene carboxyl microspheres introduce carboxyl practical groups (– COOH) on the basis of PS microspheres, making their surface efficient in further chemical combining. These carboxyl teams can be covalently adhered to nucleic acid probes, proteins or other ligands with amino teams via activation systems such as EDC/NHS, thereby attaining a lot more secure molecular addiction. Consequently, from a structural point of view, CPS microspheres have extra advantages in functionalization possibility.
Nucleic acid removal generally consists of actions such as cell lysis, nucleic acid release, nucleic acid binding to solid stage carriers, cleaning to remove pollutants and eluting target nucleic acids. In this system, microspheres play a core duty as strong stage providers. PS microspheres primarily rely on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding efficiency has to do with 60 ~ 70%, however the elution efficiency is reduced, only 40 ~ 50%. In contrast, CPS microspheres can not only use electrostatic effects however also achieve even more strong addiction via covalent bonding, reducing the loss of nucleic acids throughout the cleaning procedure. Its binding effectiveness can get to 85 ~ 95%, and the elution effectiveness is likewise enhanced to 70 ~ 80%. Furthermore, CPS microspheres are additionally significantly much better than PS microspheres in regards to anti-interference ability and reusability.
In order to confirm the performance differences in between both microspheres in actual procedure, Shanghai Lingjun Biotechnology Co., Ltd. carried out RNA removal experiments. The speculative examples were originated from HEK293 cells. After pretreatment with standard Tris-HCl buffer and proteinase K, 5 mg/mL PS and CPS microspheres were used for extraction. The results revealed that the ordinary RNA return 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 optimal value of 1.91, and the RIN worth reached 8.1. Although the procedure time of CPS microspheres is slightly longer (28 mins vs. 25 minutes) and the price is higher (28 yuan vs. 18 yuan/time), its extraction quality is substantially enhanced, and it is preferable for high-sensitivity discovery, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the viewpoint of application circumstances, PS microspheres appropriate for large screening tasks and preliminary enrichment with low needs for binding specificity due to their low cost and straightforward operation. Nevertheless, their nucleic acid binding capability is weak and conveniently influenced by salt ion concentration, making them inappropriate for long-term storage space or duplicated use. In contrast, CPS microspheres appropriate for trace sample removal because of their rich surface area practical groups, which facilitate more functionalization and can be utilized to create magnetic grain discovery packages and automated nucleic acid extraction systems. Although its prep work process is reasonably complicated and the price is fairly high, it shows stronger adaptability in clinical research study and scientific applications with strict demands on nucleic acid removal efficiency and purity.
With the quick development of molecular diagnosis, genetics modifying, liquid biopsy and other areas, greater needs are put on the effectiveness, pureness and automation of nucleic acid removal. Polystyrene carboxyl microspheres are gradually changing typical PS microspheres because of their excellent binding efficiency and functionalizable qualities, coming to be the core choice of a brand-new generation of nucleic acid extraction products. Shanghai Lingjun Biotechnology Co., Ltd. is additionally continually enhancing the particle size distribution, surface density and functionalization effectiveness of CPS microspheres and developing matching magnetic composite microsphere products to meet the needs of scientific diagnosis, clinical research study establishments and industrial customers for high-grade nucleic acid removal remedies.
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