Library construction method for DNA

Abstract

The invention discloses a library construction method for DNA. The method comprises the steps that form processing, sample preparation, reagent and consumable material preparation and instrument preparation are conducted; tail end repairing mix is added, and incubation of an incubator is subjected to tail end repairing; B1 magnetic beads are added for carrying out first-step purification, washingis carried out two times by using 75% of ethyl alcohol, and TE is dissolved to obtain a purified flat-end DNA sample; the sample is transferred to a subpackaged PCR board of a corresponding label, blowing and beating and uniform mixing are carried out, and joint connection is performed; B2 magnetic beads are added for carrying out purification, and 75% of ethyl alcohol is prepared when the sampleis put on a shelf; washing is carried out two times by using the 75% of ethyl alcohol, TE is dissolved and is transferred to the subpackaged PCR board, and blowing and beating and uniform mixing are carried out to conduct PCR; B2 magnetic beads are added for carrying out purification, when the sample is put on the shelf for 8 minutes, 75% of ethyl alcohol is prepared, washing is carried out two times by using the ethyl alcohol, and 50 TE is dissolved; a scientific and reasonable process makes connection activity be strengthened and improves the joint connecting efficiency so that more repairedDNA segments are successfully connected to a joint sequence, amplification is carried out by using a primer complemented with the joint sequence, and a sequencing library with more target amplification segments is obtained.

Description

technical field [0001] The invention belongs to the field of gene detection, and specifically refers to a DNA library construction 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 increase the speed and parallelization of analysis, and it is also difficult to reduce the cost of sequencing 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. The second-generation technology has greatly reduced the cost of sequencing, and also greatly increased the speed of sequencing while maintaining high accuracy. Among them, Illumina's first sequence...

AI Technical Summary

Problems solved by technology

[0008] The main problem to be solved in this application is to provide a DNA library construction method, which is simple to operate, scientific in process, clear in standards, and good in practice, so that its ligation activity is enhanced, adapter ligation efficiency is improved, and more repaired DNA fragments are successfully repaired. The linker sequence is connected, and then amplified by primers complementary to the linker sequence, and then a sequencing library with more target amplified fragments is obtained to solve the high cost, unscientific process, and unclear standards of a DNA library construction method. , the practical operation effect is not good, the operation is difficult, and it is difficult to copy the technical problems of the operation