{"doi":"10.1002/0471142727.mb0422s107","title":"Preparation of Single‐Cell RNA‐Seq Libraries for Next Generation Sequencing","abstract":"For the past several decades, due to technical limitations, the field of transcriptomics has focused on population-level measurements that can mask significant differences between individual cells. With the advent of single-cell RNA-Seq, it is now possible to profile the responses of individual cells at unprecedented depth and thereby uncover, transcriptome-wide, the heterogeneity that exists within these populations. This unit describes a method that merges several important technologies to produce, in high-throughput, single-cell RNA-Seq libraries. Complementary DNA (cDNA) is made from full-length mRNA transcripts using a reverse transcriptase that has terminal transferase activity. This, when combined with a second \"template-switch\" primer, allows for cDNAs to be constructed that have two universal priming sequences. Following preamplification from these common sequences, Nextera XT is used to prepare a pool of 96 uniquely indexed samples ready for Illumina sequencing.","journal":"Current Protocols in Molecular Biology","year":2014,"id":11961,"datarank":0.863361332038037,"base_score":5.755742213586912,"endowment":5.755742213586912,"self_citation_contribution":0.863361332038037,"citation_network_contribution":0.0,"self_endowment_contribution":0.863361332038037,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":315,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.1585,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2014-07-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":25301,"name":"David Gennert","orcid":"0000-0002-8668-3601","position":1,"is_corresponding":false},{"id":25302,"name":"Diana Lu","orcid":null,"position":2,"is_corresponding":false},{"id":167,"name":"Rahul Satija","orcid":"0000-0002-1479-664X","position":3,"is_corresponding":false},{"id":1413,"name":"Alex K. Shalek","orcid":"0000-0001-5670-8778","position":4,"is_corresponding":false},{"id":29633,"name":"Prisca Liberali","orcid":"0000-0003-0695-6081","position":5,"is_corresponding":false},{"id":20635,"name":"John J. Trombetta","orcid":null,"position":0,"is_corresponding":true}],"reference_count":13,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-03-01T18:20:47.508186Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}