DNA library quantifying and sequencing method

Abstract

The invention discloses a DNA library quantifying and sequencing method. The method comprises the steps that step 1, preparation of processing a sheet, a sample and a reagent is performed; step 2, a 1.5-milliliter low-adsorption tube is prepared, 499 microliter of NF water is added, and number and NF water volume are verified; a melt sample is vibrated, evenly mixed and transiently centrifuged tothe bottom of the tube, 1 microliter of sample is added into 499 microliter of NF water, tube number is verified, the diluted sample is vibrated, evenly mixed and transiently centrifuged; MIX is prepared on an ice box, 18 microliter of MIX is put into a 0.2-milliliter PCR plate placed on a metal ice box; number is verified, 2 microliter of sample is extracted and put into 18 microliter of MIX; after film sealing is carried out, centrifugation is carried out, and detection is performed in a PCR instrument; step 3, according to a quantifying detection result in step 2, an instrument-on sheet ismade, and emulsion PCR is carried out; step 4, OT instrument-off processing is carried out; and step 6, instrument-on sequencing is carried out. The DNA library quantifying and sequencing method has the advantages that the operation is simple, convenient and flexible, the sequencing cost is reduced, the sequencing processes are reduced, the labor efficiency is improved, the processes are scientific, the standard is clear, and the practical operation effect is good.

Description

technical field [0001] The invention belongs to the field of gene detection, and specifically refers to a DNA library quantification and sequencing method. Background technique [0002] The rapid acquisition of genetic information of living organisms is of great significance to the research of life sciences. The reliance of the first-generation sequencer on electrophoretic separation technology makes it difficult to further improve the analysis speed and parallelization degree, and it is also difficult to reduce the sequencing cost through miniaturization. After continuous technical development and improvement, at the beginning of the 21st century, the second-generation sequencing technology marked by Roche454 technology, Illumina's GenomeAnalyzer technology and ABI's Solid technology was born. Second-generation technology has greatly reduced the cost of sequencing, and has also greatly increased the speed of sequencing while maintaining high accuracy. Among them, Illumina...

AI Technical Summary

Problems solved by technology

[0006] The main problem to be solved by this application is to provide a DNA library quantification and sequencing method with simple operation, scientific process, clear standards, and good practical operation effect, so as to solve the high cost and unscientific process of a DNA library quantification and sequencing method. The standard is not clear, the practical operation effect is not good, the operation is difficult, and the technical problems that are difficult to replicate