The Workflow of NGS Library Preparation Kit
Celemics provides the library preparation kit for customers using Illumina, MGI, and Thermo Fisher Ion Torrent NGS platforms. There are two types of Celemics’ exclusive library preparation kits for all three platforms: LP kit and EP kit. This post aims to introduce Celemics’ products and workflow of our LP and EP kit.
- NGS Library Preparation(LP) Kit | Workflow
Celemics LP kit is suitable for the sample already fragmented by sonication or enzymatic fragmentation reaction. It is also suitable for the DNA which has an appropriate size for the short-read sequencing platform such as cell-free DNA. The detailed procedure slightly differs depending sequencing platforms, however, the fundamental workflow of Celemics LP kit is provided as below:
-
End Repair/A-tailing (ER/A): Fragmented DNA can have sticky ends. ER/A step makes those sticky ends into blunt ends and attaches the A-base in both 5’ ends. This A-base will be used at the Adapter ligation step. Please note that the A-tailing step is not required for the Ion torrent library preparation kit.
-
Adapter ligation: During this step, the adapters for each sequencing platform are attached to ER/A-treated DNA. The indices will also be attached to the samples for the MGI and Ion Torrent platform which will be used to distinguish the samples.
-
Purification: The purification step is for removing enzymes and buffers from the ER/A and Adapter ligation step.
-
PCR: The purified DNA library is amplified by PCR method for sequencing or further experiments. During this step, indices will be attached to the samples for the Illumina platform which will be used to distinguish the samples.
-
- NGS Library Preparation (EP) Kit | Workflow
Celemics EP kit is suitable for unfragmented DNA such as gDNA. It is designed to perform fragmentation, End Repair, and A-tailing in a single reaction. Like our LP kit, the detailed procedure differs depending on sequencing platforms, however, the fundamental workflow of the Celemics EP kit is provided as below:
- Fragmentation/End Repair/A-tailing (ER/A): To sequence the large DNA using a short-read NGS platform, samples should be fragmented before or during the library preparation process. Unlike the LP kit, Celemics EP kit does not require additional sonication or enzymatic fragmentation process. The EP kit allows fragmentation, End repair, and A-tailing in a single reaction tube. Please note that the A-tailing step is not required for the Ion torrent library preparation for EP kit as well.
- Adapter ligation: During this step, the adapters for each sequencing platform are attached to fER/A-treated DNA. The indices will be attached to the samples for the MGI and Ion Torrent platform which will be used to distinguish the samples.
- Purification: This is a purification step for removing enzymes and buffers from the fER/A and Adapter ligation step.
- PCR: The purified DNA library is amplified by PCR method for sequencing or further experiments. During this step, indices will be attached to the samples for the Illumina platform which will be used to distinguish the samples.
- Purification: This purification step is to remove enzyme and buffer from the PCR step.
Using Celemics LP and EP kit allow efficient yet robust generation of the NGS library within 2.5 hours regardless of the sequencing platform. For more information, please contact your local Celemics representative or contact us at supprort@celemics.com
▶ More Information
- Product page (https://www.celemics.com/products/modular-accessories/library-preparationaration-kit/)
- Video Manual