Detailed Library Preparation Steps of MGISEQ

1. Fragmentation of DNA:

The genomic DNA (gDNA) is first fragmented to obtain the desired size for sequencing.

2. End Repair and A-tailing:

The fragmented DNA undergoes end repair to create blunt ends, followed by the addition of an ‘A’ base to the 3′ ends to facilitate adapter ligation.

3. Adapter Ligation:

Specific adapters are ligated to the ends of the DNA fragments. This step is crucial for the subsequent amplification and sequencing processes.

4. PCR Amplification:

The library is then amplified using PCR to enrich the fragments that have successfully ligated adapters. This step typically focuses on amplifying the positive strand of the library , .

5. Purification:

The amplified library is purified to remove any unligated adapters and other contaminants.

6. Quality Control:

The quality and quantity of the library are assessed to ensure it meets the requirements for sequencing.

These steps collectively prepare the gDNA library for the sequencing process, ensuring that the DNA fragments are suitable for high-throughput sequencing technologies .