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-day biotechnology, microsphere products are widely utilized in the removal and purification of DNA and RNA as a result of their high particular area, excellent chemical stability and functionalized surface residential or commercial properties. Amongst them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are among both most widely examined and applied materials. This article is supplied with technical assistance and information evaluation by Shanghai Lingjun Biotechnology Co., Ltd., aiming to systematically compare the performance differences of these 2 kinds of products in the process of nucleic acid extraction, covering crucial indicators such as their physicochemical homes, surface area modification capability, binding performance and recovery rate, and illustrate their appropriate circumstances via experimental information.
Polystyrene microspheres are uniform polymer fragments polymerized from styrene monomers with good thermal stability and mechanical strength. Its surface is a non-polar structure and normally does not have energetic functional groups. Therefore, when it is directly used for nucleic acid binding, it needs to count on electrostatic adsorption or hydrophobic action for molecular addiction. Polystyrene carboxyl microspheres introduce carboxyl functional teams (– COOH) on the basis of PS microspheres, making their surface efficient in further chemical coupling. These carboxyl teams can be covalently adhered to nucleic acid probes, proteins or other ligands with amino groups through activation systems such as EDC/NHS, thus attaining more steady molecular fixation. For that reason, from a structural perspective, CPS microspheres have a lot more benefits in functionalization potential.
Nucleic acid removal typically includes steps such as cell lysis, nucleic acid launch, nucleic acid binding to strong phase service providers, cleaning to remove contaminations and eluting target nucleic acids. In this system, microspheres play a core role as strong phase carriers. PS microspheres primarily count on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding performance is about 60 ~ 70%, yet the elution effectiveness is low, just 40 ~ 50%. In contrast, CPS microspheres can not only utilize electrostatic results however also achieve even more strong addiction through covalent bonding, decreasing the loss of nucleic acids during the washing process. Its binding performance can get to 85 ~ 95%, and the elution performance is likewise increased to 70 ~ 80%. Additionally, CPS microspheres are also dramatically better than PS microspheres in regards to anti-interference ability and reusability.
In order to confirm the performance differences in between the two microspheres in real operation, Shanghai Lingjun Biotechnology Co., Ltd. carried out RNA extraction experiments. The experimental examples were originated 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 results showed that the typical RNA return extracted by PS microspheres was 85 ng/ μL, the A260/A280 proportion was 1.82, and the RIN value was 7.2, while the RNA yield of CPS microspheres was increased to 132 ng/ μL, the A260/A280 proportion was close to the optimal value of 1.91, and the RIN worth got to 8.1. Although the operation time of CPS microspheres is slightly longer (28 minutes vs. 25 minutes) and the cost is greater (28 yuan vs. 18 yuan/time), its removal top quality is considerably boosted, and it is preferable for high-sensitivity discovery, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the viewpoint of application situations, PS microspheres appropriate for massive screening jobs and preliminary enrichment with reduced needs for binding uniqueness due to their inexpensive and easy operation. However, their nucleic acid binding ability is weak and quickly influenced by salt ion focus, making them improper for lasting storage or repeated usage. In contrast, CPS microspheres appropriate for trace example removal because of their rich surface area functional teams, which help with further functionalization and can be used to build magnetic grain detection packages and automated nucleic acid removal systems. Although its preparation procedure is relatively complex and the cost is reasonably high, it shows stronger flexibility in clinical research and clinical applications with stringent needs on nucleic acid extraction efficiency and pureness.
With the rapid advancement of molecular diagnosis, gene modifying, fluid biopsy and other fields, greater demands are positioned on the effectiveness, purity and automation of nucleic acid removal. Polystyrene carboxyl microspheres are slowly changing traditional PS microspheres because of their excellent binding performance and functionalizable attributes, coming to be the core option of a new generation of nucleic acid extraction materials. Shanghai Lingjun Biotechnology Co., Ltd. is additionally continually optimizing the fragment size circulation, surface area thickness and functionalization performance of CPS microspheres and developing matching magnetic composite microsphere items to fulfill the requirements of medical diagnosis, scientific study organizations and industrial customers for top quality nucleic acid extraction remedies.
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