Nucleic acid extraction technology introduction

Nucleic acid introduction

Nucleic acid is divided into deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), among which RNA is divided into ribosomal RNA(rRNA), messenger RNA(mRNA) and transfer RNA(tRNA) according to its different functions. DNA is mainly concentrated in the nucleus, mitochondria and chloroform, while RNA is mainly distributed in the cytoplasm. As the material basis of gene expression, nucleic acid extraction plays an extremely important role in molecular biology research and clinical molecular diagnosis. The concentration and purity of nucleic acid extraction will directly affect the subsequent PCR, sequencing, vector construction, enzyme digestion and other experiments.

 Nucleic acid extraction and purification method 

① Phenol/chloroform extraction method

Phenol/chloroform extraction is a classical method for DNA extraction, which mainly uses two different organic solvents to treat the samples, dissolving DNA based nucleic acid in the water phase, lipids in the organic phase, and proteins between the two phases. This method has the advantages of low cost, high purity and good effect. The disadvantages are complicated operation and long time.

② Trizol method

Trizol method is a classical method for RNA extraction. The Trizol method is divided into aqueous phase and organic phase after centrifugation with chloroform, in which the RNA is dissolved in the aqueous phase, the aqueous phase is transferred to a new EP tube, precipitation is obtained after adding isopropanol, and then ethanol purification. This method is suitable for RNA extraction from animal tissues, cells and bacteria.

③ Centrifugal column purification method

Centrifuge column purification method can adsorb DNA specifically through special silicon matrix adsorption materials, while RNA and protein can pass smoothly, and then use high salt low PH to combine nucleic acid, low salt high PH value elution to separate and purify nucleic acid. The advantages are high purification concentration, high stability, no need for organic solvent, and low cost. The disadvantage is that it needs to be centrifuged step by step, more operation steps.

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④ Magnetic beads method

The magnetic beads method is to split the cell tissue sample through the lysate, release the nucleic acid in the sample, and then the nucleic acid molecules are specifically adsorbed on the surface of the magnetic bead, while impurities such as proteins and sugars are left in the liquid. Through the steps of cell tissue splitting, magnetic bead binding with nucleic acid, nucleic acid washing, nucleic acid elution, etc., pure nucleic acid is finally obtained. The advantages are simple operation and short time use, without the need of step centrifugation. It has low technical requirements and can realize automatic and mass operation. The specific combination of magnetic bead and nucleic acid makes the extracted nucleic acid with high concentration and purity. The disadvantage is that the current market price is relatively expensive.

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⑤ Other methods

In addition to the above four methods, there are boiling cracking, concentrated salt method, anionic detergent method, ultrasonic method and enzymatic method, etc.

 Type of nucleic acid extraction

Foregene has the world’s leading Direct PCR platform, double-column RNA Isolation platform(DNA-only + RNA only ). The main products include DNA/RNA isolation kits, PCR and Direct PCR reagents molecular lab reagents series.

① Total RNA extraction

Total RNA extraction samples include blood, cells, animal tissues, plants, viruses, etc. High purity and high concentration of total RNA can be obtained through total RNA extraction, which can be used in RT-PCR, chip analysis, in vitro translation, molecular cloning, Dot Blot and other experiments.

Foregene related RNA Isolation Kits

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Animal Total RNA Isolation Kit--Quickly and efficiently extract high-purity and high-quality total RNA from various animal tissues.

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Cell Total RNA Isolation Kit--Highly purified and high-quality total RNA can be obtained from various cultured cells in 11 minutes.

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Plant Total RNA Isolation Kit--Quickly extract high-quality total RNA from plant samples with low polysaccharide and polyphenol content.

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Viral RNA Isolation Kit--Rapidly isolate and purify viral RNA from samples such as plasma, serum, cell-free body fluids and cell culture supernatants.

② Genomic DNA extraction

Genomic DNA extraction samples include soil, feces, blood, cells, animal tissues, plants, viruses, etc. Genomic DNA extraction can be used in enzyme digestion, DNA library construction, PCR, antibody preparation, Western blot hybridization analysis, gene chip, high-throughput sequencing and other experiments.

Foregene related DNA Isolation Kits

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Animal Tissue DNA Isolation Kit--Rapid extraction and purification of genomic DNA from multiple sources, such as animal tissues, cells, etc.

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Blood DNA Midi Kit (1-5ml)--Quickly purify high-quality genomic DNA from anticoagulated blood (1-5ml).

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Buccal Swab/FTA Card DNA Isolation Kit--Quickly purify high-quality genomic DNA from buccal swab/FTA Card samples.

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Plant DNA Isolation Kit--Quickly purify and obtain high-quality genomic DNA from plant samples (including polysaccharides and polyphenol plant samples)

③ Plasmid extraction

Plasmid is a kind of circular small molecule DNA in cells, which is a common carrier for DNA recombination. The method of plasmid extraction is to remove RNA, separate plasmid from bacterial genomic DNA, and remove protein and other impurities to obtain relatively pure plasmid.

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General Plasmid Mini Kit--Quickly purify high-quality plasmid DNA from transformed bacteria for routine molecular biology experiments such as transformation and enzyme digestion

④ Other extraction types, miRNA extraction, etc.

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Animal miRNA Isolation Kit--Quickly and efficiently extract small RNA fragments of 20-200nt miRNA, siRNA, snRNA from various animal tissues and cells

 Requirements for nucleic acid extraction and purification results

① To ensure the integrity of the primary structure of nucleic acid.

② Reduce the interference of proteins, sugars, lipids and other macromolecules

③ There should be no organic solvent or high concentration of metal ions that can inhibit the enzyme in nucleic acid samples.

④ RNA and other nucleic acid contamination should be eliminated when extracting DNA, and vice versa.


Post time: Nov-24-2022