Comparative Analysis of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Concentrating On Nucleic Acid Extraction.
(LNJNbio Polystyrene Microspheres)
In the field of contemporary biotechnology, microsphere products are commonly utilized in the removal and filtration of DNA and RNA as a result of their high specific surface, excellent chemical stability and functionalized surface area homes. Amongst them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are among both most widely researched and used materials. This article is offered with technical assistance and information analysis by Shanghai Lingjun Biotechnology Co., Ltd., aiming to systematically contrast the performance differences of these two types of products in the process of nucleic acid removal, covering vital indications such as their physicochemical properties, surface adjustment ability, binding performance and healing rate, and show their appropriate situations via experimental information.
Polystyrene microspheres are homogeneous polymer bits polymerized from styrene monomers with excellent thermal stability and mechanical toughness. Its surface area is a non-polar framework and generally does not have active practical teams. Consequently, when it is straight used for nucleic acid binding, it requires to count on electrostatic adsorption or hydrophobic action for molecular addiction. Polystyrene carboxyl microspheres present carboxyl practical teams (– COOH) on the basis of PS microspheres, making their surface efficient in additional chemical coupling. These carboxyl teams can be covalently bound to nucleic acid probes, healthy proteins or other ligands with amino groups through activation systems such as EDC/NHS, thus attaining much more steady molecular fixation. Consequently, from an architectural viewpoint, CPS microspheres have more advantages in functionalization capacity.
Nucleic acid removal typically includes steps such as cell lysis, nucleic acid release, nucleic acid binding to solid phase service providers, washing to eliminate contaminations and eluting target nucleic acids. In this system, microspheres play a core role as strong phase providers. PS microspheres mainly rely upon electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding performance is about 60 ~ 70%, however the elution efficiency is reduced, only 40 ~ 50%. On the other hand, CPS microspheres can not just make use of electrostatic impacts however likewise accomplish even more strong fixation via covalent bonding, decreasing the loss of nucleic acids throughout the washing procedure. Its binding effectiveness can reach 85 ~ 95%, and the elution performance is also increased to 70 ~ 80%. In addition, CPS microspheres are likewise considerably better than PS microspheres in regards to anti-interference capability and reusability.
In order to verify the efficiency distinctions between both microspheres in real operation, Shanghai Lingjun Biotechnology Co., Ltd. performed RNA removal experiments. The speculative samples were originated from HEK293 cells. After pretreatment with common Tris-HCl barrier and proteinase K, 5 mg/mL PS and CPS microspheres were utilized for removal. The outcomes showed that the typical RNA yield extracted by PS microspheres was 85 ng/ Ī¼L, the A260/A280 ratio was 1.82, and the RIN value was 7.2, while the RNA return of CPS microspheres was increased to 132 ng/ Ī¼L, the A260/A280 ratio was close to the optimal value 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 expense is greater (28 yuan vs. 18 yuan/time), its extraction high quality is dramatically boosted, and it is better 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-scale screening jobs and initial enrichment with low demands for binding specificity due to their low cost and straightforward operation. Nevertheless, their nucleic acid binding ability is weak and quickly influenced by salt ion concentration, making them inappropriate for lasting storage or repeated use. In contrast, CPS microspheres appropriate for trace sample removal due to their rich surface area useful groups, which facilitate additional functionalization and can be used to create magnetic bead detection sets and automated nucleic acid removal platforms. Although its preparation process is reasonably complex and the cost is reasonably high, it shows more powerful flexibility in scientific research study and medical applications with rigorous requirements on nucleic acid removal efficiency and pureness.
With the quick growth of molecular medical diagnosis, genetics editing, fluid biopsy and various other areas, higher needs are placed on the efficiency, purity and automation of nucleic acid removal. Polystyrene carboxyl microspheres are gradually changing conventional PS microspheres as a result of their exceptional binding performance and functionalizable characteristics, becoming the core choice of a new generation of nucleic acid removal materials. Shanghai Lingjun Biotechnology Co., Ltd. is also continuously enhancing the fragment size distribution, surface density and functionalization efficiency of CPS microspheres and creating matching magnetic composite microsphere products to meet the needs of medical diagnosis, scientific research study institutions and commercial customers for top notch nucleic acid extraction solutions.
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