Analysis of Four Conventional Nucleic Acid Purification Methods

Analysis of Four Conventional Nucleic Acid Purification Methods

From the development history of nucleic acid extraction technology, nucleic acid extraction has gone through four main development stages.

• Precipitation method with organic solvent extraction technology.
• Spin column method with silica gel membrane adsorption technology.
• Magnetic bead method with magnetic bead adsorption technology.
• Resin method with ion-exchange resin adsorption technology.

Precipitation

Advantages: flexible processing volume, relatively cheap.

Disadvantages: phenol, chloroform and other reagents are more toxic and contamination is more serious. The experimental operation is demanding, the recovery rate of nucleic acid is low, and it is difficult to carry out microscopic operation, which is basically eliminated in practical application.

Spin column method

Advantages: easy and fast operation process, higher purity of nucleic acids and less inhibitors than traditional organic solvent extraction, relatively cheap.

Disadvantages: some reagents are toxic. The loading capacity of the silica column is limited, which is a limit to the initial sample size. When too much sample is put in (e.g. large pieces of tissue) it can lead to clogging of the adsorption film and thus risk of low yield or contamination.

Magnetic bead method.

Advantages: safe, non-toxic, simple operation, easy automation, and high yield of microsamples. Automated working system makes the extraction mode from single sample gradually develop to multi-sample unified nucleic acid extraction, which can realize high throughput operation, one of the future development trend.

Disadvantages: relatively expensive.

Ion-exchange resin method

Advantages: good integrity of extracted nucleic acids, high purity, suitable for large molecular nucleic acid extraction, and Single Molecule Real-Time（SMRT）Sequencing.

Disadvantages: not suitable for extraction of small fragment nucleic acid.

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