Nucleic Acid Isolation : A critical step in the field of Molecular Biology


Nucleic Acid Isolation : A critical step in the field of Molecular Biology



Effective isolation of nucleic acids (NAs) is important for clinical molecular methods, including polymerase chain reaction (PCR) and sequencing. Techniques involve sample exposure to chemicals, enzymes, or binding matrices that reduce sample volume, variability, and complexity to achieve purity goals. The isolated NA should be efficiently separated from inhibitors of a downstream molecular assay. The best preparation method depends on the requirements for a particular application. Goals may include flexibility for multiple sample types, large batch processing, speed, or high-purity NA. In today’s molecular biology studies better results can usually be obtained by the right combination of lysis, concentration, purification, and efficiency for NA isolation. NA preparation techniques have improved significantly over the past 60 years in speed and complexity.

Nucleic acid preparation can refer to isolation, extraction, purification, and/or separation. Three steps are used for most samples: (1) extracting or releasing the NA, typically from cells; (2) separating or isolating NA from other material; and (3) purifying the NA by removal of inhibitory substances. Concentrating NA can also be important for the detection of low-level analytes.

How to Extract Nucleic Acids – Process & Methods

There are various technologies for the extraction of nucleic acids: Chemical method, Magnetic bead technology and Spin column technology.Early isolations of NA used tedious methods to isolate NA away from other cellular components, often including alkaline lysis followed by acid and alcohol precipitation (Chemical method).

Spin column-based NA purification is a solid phase extraction method to quickly purify nucleic acids.Sorption processes based on (a) hydrogen-binding interaction with an underivatised hydrophilic matrix, typically silica, under chaotropic conditions, (b) ionic exchange under aqueous conditions by means of an anion exchanger, (c) affinity and (d) size exclusion mechanisms were used for DNA purification. Solid-phase systems which adsorb DNA—silica-based particles, glass fibres, and anion-exchange carriers–are used in chromatographic separation columns.

Magnetic separation is an emerging technology that uses magnetism for the efficient separation of micrometre-sized para- and ferromagnetic particles from chemical or biological suspensions. The first magnetic particle used for extraction consisted of an iron-oxide core covered by functional carboxylic group, which then binds DNA or RNA Since then, many modifications have been made using different surface functional group, such as sulphate, amino, and hydroxyl groups. Besides these functional groups, preactivated magnetic beads with different functional groups are available such as tosyl or epoxy groups. Magnetic beads activated with protein A, protein G, or streptavidin are commercially available. Magnetic bead separation presents many benefits over centrifuge-dependent extraction process by allowing an equipment-free process. Magnetic separation of NA has several advantages compared to other techniques used for the same purpose. NA can be isolated directly from crude sample materials such as blood, tissue homogenates, cultivation media, water, etc. The particles are used in batch processes where there are hardly any restrictions with respect to the sample volumes

BTL Biotechno Labs Pvt Ltd is the best supplier of NA isolation kits from Omega Biotek. Their Mag-Bind® magnetic bead-based systems consist of 7 types of magnetic beads. Omega Biotek matches our magnetic beads with optimized buffers to achieve optimal purification efficiency. Our kits support the following liquid handlers and magnetic processers:

  • Hamilton Microlab® Prep™
  • Hamilton Microlab® STAR™
  • Hamilton Microlab® STARlet™
  • Hamilton Microlab® NIMBUS™
  • Beckman Coulter Biomek® FX
  • Beckman Coulter Biomek® NX
  • Tecan EVO
  • Tecan Fluent® Automation Workstation
  • Opentron OT-2
  • ThermoKingFisher® Flex
  • ThermoKingFisher® DUO
  • QiagenBioSprint® 96
  • Applied BiosystemsMagMAX® 96

We supply DNA and RNA isolation kits from Blood, Stool, cells, tissues, soil, water, pathogen: virus and bacteria, plant and fungus and many more. Further the company also provides DNA/RNA clean up kit, Gel extraction kit, PCR clean up kit, Plasmid isolation kit. Not only this the company provides PCR reagents, Electrophoresis buffers, other related reagents and plastic wares.