Relative Analysis 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-day biotechnology, microsphere products are commonly made use of in the removal and filtration of DNA and RNA because of their high details area, excellent chemical security and functionalized surface residential or commercial properties. Among them, polystyrene (PS) microspheres and their acquired polystyrene carboxyl (CPS) microspheres are just one of the two most extensively researched and applied products. This article is provided with technological support and data analysis by Shanghai Lingjun Biotechnology Co., Ltd., aiming to systematically compare the efficiency differences of these 2 types of products in the procedure of nucleic acid removal, covering key indications such as their physicochemical residential properties, surface alteration ability, binding performance and healing price, and illustrate their relevant situations via experimental data.
Polystyrene microspheres are homogeneous polymer fragments polymerized from styrene monomers with great thermal stability and mechanical stamina. Its surface area is a non-polar structure and typically does not have active functional teams. Consequently, when it is directly used for nucleic acid binding, it requires to rely on electrostatic adsorption or hydrophobic activity for molecular addiction. Polystyrene carboxyl microspheres introduce carboxyl functional teams (– COOH) on the basis of PS microspheres, making their surface area capable of more chemical combining. These carboxyl groups can be covalently adhered to nucleic acid probes, proteins or various other ligands with amino teams via activation systems such as EDC/NHS, thus attaining a lot more stable molecular fixation. As a result, from an architectural viewpoint, CPS microspheres have a lot more benefits in functionalization capacity.
Nucleic acid extraction normally consists of steps such as cell lysis, nucleic acid launch, nucleic acid binding to solid stage carriers, cleaning to eliminate pollutants and eluting target nucleic acids. In this system, microspheres play a core function as strong stage providers. PS microspheres mostly rely on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding performance is about 60 ~ 70%, however the elution efficiency is low, only 40 ~ 50%. On the other hand, CPS microspheres can not only utilize electrostatic impacts however likewise attain even more solid fixation with covalent bonding, minimizing the loss of nucleic acids during the washing process. Its binding performance can reach 85 ~ 95%, and the elution performance is likewise raised to 70 ~ 80%. On top of that, CPS microspheres are likewise substantially much better than PS microspheres in terms of anti-interference capability and reusability.
In order to validate the performance differences in between the two microspheres in real operation, Shanghai Lingjun Biotechnology Co., Ltd. performed RNA extraction experiments. The experimental samples were stemmed 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 removal. The outcomes showed that the typical RNA yield removed by PS microspheres was 85 ng/ Ī¼L, 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/ Ī¼L, the A260/A280 ratio was close to the perfect worth of 1.91, and the RIN value reached 8.1. Although the operation 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 top quality is substantially enhanced, and it is preferable 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 appropriate for massive screening tasks and initial enrichment with reduced demands for binding specificity because of their low cost and easy procedure. However, their nucleic acid binding capability is weak and quickly affected by salt ion focus, making them inappropriate for long-term storage or repeated use. On the other hand, CPS microspheres are suitable for trace sample removal as a result of their rich surface area useful groups, which promote more functionalization and can be made use of to create magnetic grain discovery kits and automated nucleic acid extraction systems. Although its preparation process is relatively intricate and the cost is fairly high, it shows stronger versatility in scientific research study and clinical applications with stringent needs on nucleic acid removal effectiveness and pureness.
With the quick advancement of molecular medical diagnosis, gene editing, fluid biopsy and various other fields, greater demands are positioned on the performance, purity and automation of nucleic acid extraction. Polystyrene carboxyl microspheres are gradually changing conventional PS microspheres due to their superb binding efficiency and functionalizable features, becoming the core selection of a new generation of nucleic acid removal materials. Shanghai Lingjun Biotechnology Co., Ltd. is additionally constantly maximizing the fragment size distribution, surface area thickness and functionalization effectiveness of CPS microspheres and creating matching magnetic composite microsphere products to meet the needs of clinical medical diagnosis, scientific study institutions and commercial customers for premium nucleic acid removal remedies.
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