Automated nucleic acid extraction, detection and quantification platform

In the field of molecular biology, the nucleic acid extraction, detection and quantification of samples is a common experimental method in every laboratory. In recent years, the birth of real-time quantitative PCR and second-generation sequencing technology has greatly increased the demand for nucleic acid extraction. These new nucleic acid detection technologies require relatively high purity of nucleic acids, so the quality of nucleic acid extraction is more important than before. On the other hand, accurate nucleic acid quantification plays a key role in the success or failure of these new technologies, such as second-generation sequencing. Therefore, today's nucleic acid extraction, detection and quantification technologies have undergone major changes.

Nucleic acid extraction

In the past, nucleic acid extraction was carried out by using a human hand with a kit. Since it is manually operated, it often takes a long time when the flux is high. Moreover, based on the experience and proficiency of different experimental personnel, there may be differences in the consistency of time and purity of nucleic acid extraction. Faced with these problems, the process of nucleic acid extraction began to move toward automation to ensure the stability of the extraction effect. Among the classical nucleic acid extraction and purification methods, the biomagnetic beads method is the best. The technology coats the purification medium on the surface of the nano-scale biomagnetic beads, and adsorbs the nucleic acid through the medium, and attaches the nucleic acid to the magnetic beads to move in an external magnetic field, thereby achieving the function of solid-liquid separation and purification. Therefore, to achieve automated nucleic acid extraction, the magnetic bead method is the best choice. Because the magnetic bead method can be performed entirely on the pipetting platform, and the extraction purity and yield are high ( 500 μg of DNA per mg of magnetic beads can be adsorbed ) , and 96 or 384 sample nucleic acid extractions can be performed simultaneously . This approach has begun to receive attention in high-throughput laboratories. There are many magnetic bead extraction nucleic acid kits available on the market, most of which can be combined with pipetting platforms.

Nucleic acid detection and quantification

After nucleic acid extraction, the next step is to detect and quantify. This step is very important for downstream second-generation sequencing experiments. If the quantification is inaccurate, it will greatly affect the results of sequencing, resulting in wasted high experimental costs. The traditional nucleic acid detection uses the ultraviolet spectroscopic brightness method to determine whether there is protein contamination by the ratio of A260 to A280 . In TE buffer, the A260/A280 ratio of pure DNA was 1.8 , and the A260/A280 ratio of pure RNA was 2.0 . Both the increase and decrease in the ratio indicate impure. Both the protein and the phenol added in the nucleic acid extraction reduce the ratio. The high absorption peak of the protein at 280 nm and the phenol at 270 nm can identify whether it is mainly protein contamination or phenol contamination. The contamination of RNA can cause the ratio of DNA products to be higher than 1.8 . Therefore , the DNA solution with a ratio of 1.8 is not necessarily a pure DNA solution, and may be contaminated with protein, phenol and RNA , and needs to be identified by other methods. More accurate methods include fluorescence quantitative PCR and fluorescent staining, such as PicoGreen , EB , Oligreen, etc., among which PicoGreen fluorescence method is most convenient and accurate. PicoGreen is one of the more commonly used fluorescent dyes for nucleic acid quantification. It binds to double-stranded DNA and has very good specificity. PicoGreen can be combined with DNA for high-throughput assays using a fluorescent microplate reader.

In order to solve the problems of flux and consistency, nucleic acid extraction and detection require a more efficient and accurate platform. Boqi Technology also introduced the high-throughput automatic pipetting workstation of the American pipetting platform pioneer, Apricot Desings , and the German BMG Labtech multi-function microplate reader, and combined to become an automated nucleic acid extraction, detection and quantification platform. The TPS series from Apricot Designs in the United States is a versatile pipetting platform that not only enables automated pipetting but also achieves very good accuracy. Automated and high-throughput nucleic acid extraction with the magnetic bead kit on the TPS platform enables up to 480 samples of nucleic acid extraction and purification in up to 40 minutes . In addition to nucleic acid extraction, the TPS system can perform a variety of experiments, such as PCR , plate-to-plate pipetting, gradient dilution, ELISA , and more. Therefore, it is ideal for high-throughput and diverse research units.

Therefore, the combination of Apricot Designs ' TPS nucleic acid extractor and BMG Labtech FLUOstar Omega multi- plate reader, Biotech can provide a comprehensive, stable, accurate and automated nucleic acid extraction, detection and quantification platform. Combined with these two systems, high-throughput nucleic acid extraction, purification, and assay quantification can be easily and simply accomplished. So far, many research units and large-scale sequencing organizations have used our platform to solve the problem of nucleic acid extraction and detection. This is also a major trend in high-throughput molecular biology research!

Reference materials :

Http://bmglabtech.customers.condeon.net/media/35216/1044002.pdf

Http://bmglabtech.customers.condeon.net/media/35216/1043726.pdf

Http://bmglabtech.customers.condeon.net/media/35216/1066012.pdf

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