Relative Evaluation of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Concentrating On Nucleic Acid Removal.
(LNJNbio Polystyrene Microspheres)
In the field of modern-day biotechnology, microsphere materials are extensively used in the removal and purification of DNA and RNA as a result of their high details surface, excellent chemical security and functionalized surface area residential or commercial properties. Amongst them, polystyrene (PS) microspheres and their acquired polystyrene carboxyl (CPS) microspheres are one of the two most widely examined and applied materials. This article is given with technical support and data analysis by Shanghai Lingjun Biotechnology Co., Ltd., intending to systematically compare the performance differences of these 2 types of products in the process of nucleic acid removal, covering crucial indicators such as their physicochemical properties, surface area alteration capability, binding performance and healing price, and show their applicable circumstances via speculative data.
Polystyrene microspheres are homogeneous polymer bits polymerized from styrene monomers with excellent thermal stability and mechanical stamina. Its surface is a non-polar structure and usually does not have active useful teams. Consequently, when it is directly used for nucleic acid binding, it needs to count on electrostatic adsorption or hydrophobic activity for molecular fixation. Polystyrene carboxyl microspheres present carboxyl functional 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, proteins or various other ligands with amino groups with activation systems such as EDC/NHS, consequently achieving much more steady molecular fixation. As a result, from an architectural point of view, CPS microspheres have much more advantages in functionalization capacity.
Nucleic acid extraction usually consists of steps such as cell lysis, nucleic acid release, nucleic acid binding to strong phase carriers, washing to get rid of impurities and eluting target nucleic acids. In this system, microspheres play a core duty as strong phase carriers. PS microspheres mostly depend 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 low, only 40 ~ 50%. In contrast, CPS microspheres can not only use electrostatic results however likewise attain even more strong addiction via covalent bonding, decreasing the loss of nucleic acids during the washing procedure. Its binding performance can reach 85 ~ 95%, and the elution performance is likewise increased to 70 ~ 80%. In addition, CPS microspheres are additionally considerably much better than PS microspheres in terms of anti-interference ability and reusability.
In order to confirm the performance distinctions between the two microspheres in actual procedure, Shanghai Lingjun Biotechnology Co., Ltd. conducted RNA extraction experiments. The experimental examples were stemmed from HEK293 cells. After pretreatment with typical Tris-HCl buffer and proteinase K, 5 mg/mL PS and CPS microspheres were used for extraction. The results showed that the ordinary RNA yield drawn out by PS microspheres was 85 ng/ Ī¼L, the A260/A280 proportion was 1.82, and the RIN value was 7.2, while the RNA yield of CPS microspheres was enhanced to 132 ng/ Ī¼L, the A260/A280 ratio was close to the suitable 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 expense is higher (28 yuan vs. 18 yuan/time), its extraction top quality is considerably improved, and it is better for high-sensitivity detection, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the viewpoint of application circumstances, PS microspheres are suitable for large-scale screening tasks and initial enrichment with reduced requirements for binding uniqueness due to their inexpensive and easy operation. Nevertheless, their nucleic acid binding capability is weak and easily impacted by salt ion focus, making them improper for long-term storage space or repeated usage. On the other hand, CPS microspheres are suitable for trace sample removal as a result of their abundant surface area practical teams, which facilitate further functionalization and can be used to build magnetic bead discovery kits and automated nucleic acid removal systems. Although its preparation procedure is reasonably intricate and the price is fairly high, it shows stronger flexibility in clinical research study and clinical applications with strict demands on nucleic acid extraction efficiency and purity.
With the rapid growth of molecular medical diagnosis, genetics modifying, fluid biopsy and various other fields, greater requirements are placed on the efficiency, pureness and automation of nucleic acid removal. Polystyrene carboxyl microspheres are progressively replacing traditional PS microspheres as a result of their excellent binding efficiency and functionalizable qualities, ending up being the core choice of a brand-new generation of nucleic acid extraction products. Shanghai Lingjun Biotechnology Co., Ltd. is also continually optimizing the fragment dimension circulation, surface thickness and functionalization performance of CPS microspheres and creating matching magnetic composite microsphere products to meet the needs of clinical medical diagnosis, scientific research study establishments and industrial consumers for high-grade nucleic acid extraction options.
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