Nucleic acid mass spectrometry

Nucleic acid mass spectrometry is a multiplex PCR analysis and detection system based on MALDI-TOF MS mass spectrometry. It primarily achieves nucleic  acid detection through multiplex PCR high-throughput chip time-of-flight mass spectrometry, enabling the simultaneous detection of dozens or even hundreds of targets in a single sample. moreover, the system can process over 1,000 cases per day.

The main steps of nucleic acid mass spectrometry:

1. Polymerase chain reaction (PCR): one upstream and one downstream primer is designed according to the site to be detected, and PCR is performed using sample DNA as a template to obtain a PCR product fragment containing the target site.

2. Shrimp alkaline phosphatase (SAP) treatment: SAP(10322ES) was added to dephosphorylate the PCR product to eliminate the remaining dNTP in the reaction system.

3. Single base extension reaction: Add a single base extension primer and ddNTP, single base extension reaction.

4. Resin purification: desalting resin is added to desalting and purifying the extension product to avoid the interference of salt ion peak in mass spectrometry.

5. Combination of sample and matrix: the sample is spotted on the chip with matrix for mass spectrometry detection, and different gene fragments finally form different detection peak maps.

In recent years, nucleic acid mass spectrometry has become a new platform for molecular diagnosis after PCR and NGS. Fluorescence quantitative PCR detection speed is fast, but the flux is limited, can not easily and quickly meet the detection needs of dozens to hundreds of sites for multiple genes; and high-throughput sequencing, although the flux is extremely high, but its detection cost is high, the project detection cycle is long, the detection data also need professional analysis and interpretation, the technical threshold is high; the stable and accurate detection results and low detection cost of nucleic acid mass spectrometry meet the needs of qualitative and quantitative detection of medium-flux site SNP and gene mutation, and can carry out convenient and fast DNA methylation and copy number variation (CNV) detection, which highlights its strong competitiveness in the field of gene detection, thus becoming one of the main platforms for the rapid development of life science, especially in the field of clinical diagnosis.

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