With the continuous breakthrough of high-throughput sequencing technology, the application of second-generation sequencing is now very extensive. However, the construction of NGS libraries, especially the construction of the target area capture library, is more complicated, the general process is 2-3 days, and the manual operation steps are as many as hundreds of steps. Any step of operation errors will affect the final result.
In order to improve the efficiency and throughput of capturing and constructing libraries in the target area, and reduce the uncertainty caused by manual operation, AMTK and Wuhan KaiDeWeiSi Biotechnology Co., Ltd. jointly developed the NGS hybridization elution automated process, which is one-time The throughput is 96 samples. This automated process is combined with the pathogen liquid phase hybridization capture kit independently developed by KaiDeWeiSi to hybridize and capture DNA from human epithelial cells to construct a library to diagnose specific pathogen infections and provide decision-making basis for clinical treatment.
Automation solution
Next-generation sequencing hybridization elution flow chart
In the LH-1209 high-throughput liquid processing workstation, the hybridization elution process of 91 samples was automatically completed
Step 1: Open the software and select the "hybridization elution" program
Step 2: According to the requirements of the deck, carry out the deck layout, as shown in the figure below
Step 3: Click "Start" to run, the program runs
Experimental results and data display
Sequencing quality
Effective Reads ratio
Remarks: Effective Reads=Human&Target Pathogen&Control
Sample typing results
Average sequencing depth of some samples
From the point of view of the experimental procedures, the LH-1209 liquid workstation workstation can help users carry out the whole process of NGS hybridization and elution, with a single-stage layout and easy operation. Judging from the experimental results, the data of 91 samples meet the quality control standards of bio-information analysis and are reliable.
The realization of the automated process of hybridization elution improves the efficiency of next-generation sequencing library construction and promotes the clinical application of second-generation sequencing technology.