Relative Evaluation 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 biotechnology, microsphere materials are commonly utilized in the extraction and purification of DNA and RNA due to their high certain area, good chemical stability and functionalized surface area residential properties. Among them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are among both most widely researched and used products. This short article is provided with technical assistance and information analysis by Shanghai Lingjun Biotechnology Co., Ltd., intending to systematically contrast the efficiency differences of these 2 sorts of materials in the process of nucleic acid extraction, covering key indications such as their physicochemical buildings, surface area alteration ability, binding effectiveness and recovery rate, and highlight their applicable situations through speculative data.
Polystyrene microspheres are uniform polymer bits polymerized from styrene monomers with great thermal stability and mechanical strength. Its surface is a non-polar structure and typically does not have active functional groups. Therefore, when it is directly used for nucleic acid binding, it requires to count on electrostatic adsorption or hydrophobic activity for molecular addiction. Polystyrene carboxyl microspheres introduce carboxyl useful groups (– COOH) on the basis of PS microspheres, making their surface area capable of further chemical coupling. These carboxyl teams can be covalently bound to nucleic acid probes, proteins or various other ligands with amino teams via activation systems such as EDC/NHS, thereby attaining more steady molecular fixation. Consequently, from a structural perspective, CPS microspheres have more benefits in functionalization potential.
Nucleic acid removal typically consists of steps such as cell lysis, nucleic acid release, nucleic acid binding to solid stage service providers, cleaning to get rid of contaminations and eluting target nucleic acids. In this system, microspheres play a core role as solid phase carriers. PS microspheres mainly depend on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding effectiveness is about 60 ~ 70%, but the elution efficiency is low, just 40 ~ 50%. In contrast, CPS microspheres can not only utilize electrostatic effects yet additionally achieve even more strong addiction with covalent bonding, reducing the loss of nucleic acids throughout the cleaning process. Its binding effectiveness can reach 85 ~ 95%, and the elution efficiency is additionally boosted to 70 ~ 80%. In addition, CPS microspheres are also dramatically much better than PS microspheres in terms of anti-interference capability and reusability.
In order to confirm the efficiency distinctions in between the two microspheres in real operation, Shanghai Lingjun Biotechnology Co., Ltd. conducted RNA removal experiments. The experimental samples were derived from HEK293 cells. After pretreatment with standard Tris-HCl buffer and proteinase K, 5 mg/mL PS and CPS microspheres were used for removal. The outcomes revealed that the average RNA yield removed 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 boosted to 132 ng/ μL, the A260/A280 proportion was close to the ideal value of 1.91, and the RIN worth reached 8.1. Although the operation time of CPS microspheres is a little longer (28 minutes 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 situations, PS microspheres appropriate for massive screening projects and preliminary enrichment with low demands for binding specificity due to their inexpensive and straightforward operation. However, their nucleic acid binding capability is weak and easily affected by salt ion focus, making them improper for lasting storage space or duplicated use. On the other hand, CPS microspheres appropriate for trace example removal due to their rich surface useful groups, which promote additional functionalization and can be utilized to create magnetic bead detection kits and automated nucleic acid removal platforms. Although its prep work process is fairly intricate and the expense is reasonably high, it reveals stronger flexibility in scientific research and medical applications with rigorous needs on nucleic acid extraction effectiveness and purity.
With the fast advancement of molecular diagnosis, gene modifying, fluid biopsy and other fields, higher needs are placed on the effectiveness, pureness and automation of nucleic acid removal. Polystyrene carboxyl microspheres are slowly replacing conventional PS microspheres because of their excellent binding efficiency and functionalizable attributes, coming to be the core choice of a new generation of nucleic acid extraction products. Shanghai Lingjun Biotechnology Co., Ltd. is likewise constantly optimizing the fragment size distribution, surface thickness and functionalization efficiency of CPS microspheres and establishing matching magnetic composite microsphere items to meet the needs of professional diagnosis, scientific study organizations and commercial consumers for high-quality nucleic acid extraction services.
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