<\/figure>\n\n\n\n1. 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":"\n
NGS\u81ea\u52a8\u5316\u5efa\u5e93\u7684\u6d41\u7a0b - Opentrons\u4e2d\u56fd\u5b98\u7f51<\/title>\n \n \n \n \n \n \n \n \n \n \n \n\t \n\t \n\t \n \n \n \n\t \n\t \n\t \n