Comparative Analysis of the Application of Polystyrene Microspheres and Polystyrene Carboxyl Microspheres in Biotechnology – Focusing on Nucleic Acid Removal.
(LNJNbio Polystyrene Microspheres)
In the field of contemporary biotechnology, microsphere materials are extensively made use of in the removal and filtration of DNA and RNA due to their high certain surface, excellent chemical stability and functionalized surface residential properties. Amongst them, polystyrene (PS) microspheres and their obtained polystyrene carboxyl (CPS) microspheres are one of the two most extensively studied and applied materials. This post is provided with technological assistance and information analysis by Shanghai Lingjun Biotechnology Co., Ltd., intending to systematically contrast the performance differences of these 2 types of products in the procedure of nucleic acid extraction, covering vital indicators such as their physicochemical residential or commercial properties, surface area alteration capability, binding performance and recovery rate, and illustrate their suitable situations via experimental information.
Polystyrene microspheres are uniform polymer particles polymerized from styrene monomers with good thermal stability and mechanical strength. Its surface area is a non-polar structure and typically does not have energetic useful teams. For that reason, when it is directly made use of for nucleic acid binding, it needs to count on electrostatic adsorption or hydrophobic activity for molecular fixation. Polystyrene carboxyl microspheres present carboxyl useful teams (– COOH) on the basis of PS microspheres, making their surface area efficient in more chemical combining. These carboxyl groups can be covalently bonded to nucleic acid probes, healthy proteins or various other ligands with amino groups through activation systems such as EDC/NHS, therefore achieving a lot more secure molecular addiction. Therefore, from an architectural viewpoint, CPS microspheres have extra benefits in functionalization potential.
Nucleic acid extraction normally consists of steps such as cell lysis, nucleic acid launch, nucleic acid binding to solid stage carriers, washing to get rid of contaminations and eluting target nucleic acids. In this system, microspheres play a core function as solid stage carriers. PS microspheres primarily count on electrostatic adsorption and hydrogen bonding to bind nucleic acids, and their binding efficiency has to do with 60 ~ 70%, but the elution performance is low, only 40 ~ 50%. On the other hand, CPS microspheres can not only utilize electrostatic effects but likewise achieve more strong addiction via covalent bonding, lowering the loss of nucleic acids throughout the washing process. Its binding effectiveness can reach 85 ~ 95%, and the elution effectiveness is also raised to 70 ~ 80%. Additionally, CPS microspheres are likewise considerably better than PS microspheres in regards to anti-interference capability and reusability.
In order to validate the performance distinctions in between both microspheres in real procedure, Shanghai Lingjun Biotechnology Co., Ltd. performed RNA removal experiments. The speculative samples were originated from HEK293 cells. After pretreatment with standard Tris-HCl barrier and proteinase K, 5 mg/mL PS and CPS microspheres were used for extraction. The results revealed that the average RNA return drawn out 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 enhanced to 132 ng/ μL, the A260/A280 proportion was close to the optimal value of 1.91, and the RIN value reached 8.1. Although the operation time of CPS microspheres is somewhat longer (28 minutes vs. 25 mins) and the expense is higher (28 yuan vs. 18 yuan/time), its extraction quality is substantially enhanced, and it is more suitable for high-sensitivity discovery, such as qPCR and RNA-seq.
( SEM of LNJNbio Polystyrene Microspheres)
From the perspective of application scenarios, PS microspheres appropriate for large screening jobs and preliminary enrichment with reduced requirements for binding uniqueness because of their inexpensive and straightforward procedure. Nevertheless, their nucleic acid binding capacity is weak and quickly influenced by salt ion concentration, making them unsuitable for lasting storage or repeated usage. In contrast, CPS microspheres appropriate for trace sample removal due to their abundant surface area useful groups, which assist in more functionalization and can be made use of to create magnetic bead discovery sets and automated nucleic acid removal systems. Although its preparation process is relatively complex and the price is relatively high, it reveals more powerful flexibility in clinical study and medical applications with strict demands on nucleic acid extraction efficiency and pureness.
With the quick development of molecular medical diagnosis, genetics editing and enhancing, liquid biopsy and various other areas, higher requirements are positioned on the effectiveness, pureness and automation of nucleic acid removal. Polystyrene carboxyl microspheres are slowly replacing conventional PS microspheres because of their superb binding performance and functionalizable characteristics, ending up being the core choice of a new generation of nucleic acid removal products. Shanghai Lingjun Biotechnology Co., Ltd. is additionally continually optimizing the particle dimension distribution, surface area thickness and functionalization performance of CPS microspheres and developing matching magnetic composite microsphere items to meet the requirements of professional medical diagnosis, scientific research organizations and industrial customers for premium nucleic acid removal options.
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