{"id":3548,"date":"2024-07-27T17:59:37","date_gmt":"2024-07-27T09:59:37","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&p=3548"},"modified":"2024-09-02T18:21:36","modified_gmt":"2024-09-02T10:21:36","slug":"ngs-jiankuliucheng","status":"publish","type":"post","link":"https:\/\/en.opentrons.com.cn\/news\/ngs-jiankuliucheng\/","title":{"rendered":"NGS automated library construction process"},"content":{"rendered":"\n

NGS automated library construction refers to the use of automated technology and equipment to carry out a high-throughput, high-efficiency, and high-precision library construction process for DNA or RNA samples. This process covers many aspects from sample processing, DNA\/RNA extraction, library construction to quality control, etc., aiming to reduce manual operations and improve experimental efficiency and accuracy.<\/p>\n\n\n\n

\"NGS\u81ea\u52a8\u5316\u5efa\u5e93\"<\/figure>\n\n\n\n

1. Sample preparation and DNA\/RNA extraction<\/p>\n\n\n\n

1. Sample collection: Collect target samples, such as cells, tissues, blood, etc., to ensure the integrity and representativeness of the samples.<\/p>\n\n\n\n

2. Sample processing: Select the appropriate processing method according to the sample type, such as cell lysis, tissue grinding, etc., to release DNA or RNA.<\/p>\n\n\n\n

3. DNA\/RNA extraction: Use appropriate extraction methods (such as phenol-chloroform method, magnetic bead method, column method, etc.) to purify high-quality DNA or RNA from the sample.<\/p>\n\n\n\n

2. Library construction<\/p>\n\n\n\n

Library construction is the core step of automated NGS library construction, which usually includes the following sub-steps:<\/p>\n\n\n\n

1. DNA\/RNA fragmentation:<\/p>\n\n\n\n

1>Cut DNA or RNA molecules into smaller fragments for subsequent sequencing reactions. Fragmentation can be achieved by physical methods (such as sonication) or chemical methods.<\/p>\n\n\n\n

2>Fragment size is usually between 100-1000 base pairs, depending on the sequencing platform and experimental needs.<\/p>\n\n\n\n

2. End repair and A-tail addition:<\/p>\n\n\n\n

1>Perform end repair on fragmented DNA or RNA to remove end damage and add phosphate groups and 3\u2019 end A tails. These repair steps facilitate subsequent adapter ligation and library amplification.<\/p>\n\n\n\n

3. Connect the adapter:<\/p>\n\n\n\n

1>Connect the repaired DNA or RNA fragment to a specific adapter (also called an adapter). Aptamers are DNA or RNA fragments containing specific sequences that are used in subsequent PCR amplification and sequencing reactions.<\/p>\n\n\n\n

2>In the process of automated library construction, this step is usually achieved through automated pipetting and ligation reactions.<\/p>\n\n\n\n

4. Library amplification:<\/p>\n\n\n\n

1>Amplify the DNA or RNA fragments connected to the aptamer through PCR amplification to a sufficient amount to form a library that can be used for sequencing.<\/p>\n\n\n\n

2> During the amplification process, primers with adapter sequences are used for specific amplification.<\/p>\n\n\n\n

3. Library quality control<\/p>\n\n\n\n

Library quality control is an important step to ensure library quality and reliability of sequencing results. Commonly used library quality control methods include:<\/p>\n\n\n\n

1. Quantitative PCR: used to detect the concentration and amplification efficiency of the library.<\/p>\n\n\n\n

2. Polyacrylamide gel electrophoresis: used to evaluate the size distribution of library fragments.<\/p>\n\n\n\n

3. Mass spectrometry analysis: used to further verify the quality and purity of the library.<\/p>\n\n\n\n

4. Function:<\/p>\n\n\n\n

1. Automated operation: Automate sample processing, library construction and other steps through automated equipment to reduce manual intervention.<\/p>\n\n\n\n

2. Improve efficiency: The automated process shortens the experimental cycle and improves experimental efficiency.<\/p>\n\n\n\n

3. Reduce errors: Reduce errors caused by human operations and improve the accuracy and stability of experimental results.<\/p>\n","protected":false},"excerpt":{"rendered":"

NGS automated library construction refers to the use of […]<\/p>\n","protected":false},"author":5,"featured_media":3550,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"inline_featured_image":false,"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"footnotes":""},"categories":[],"tags":[],"class_list":["post-3548","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry"],"yoast_head":"\nNGS\u81ea\u52a8\u5316\u5efa\u5e93\u7684\u6d41\u7a0b - Opentrons\u4e2d\u56fd\u5b98\u7f51<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/en.opentrons.com.cn\/news\/ngs-jiankuliucheng\/\" \/>\n<meta property=\"og:locale\" content=\"zh_CN\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"NGS\u81ea\u52a8\u5316\u5efa\u5e93\u7684\u6d41\u7a0b - 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