{"doi":"10.20944/preprints202412.1876.v1","title":"Comparison of Commercially Available Thermostable DNA Polymerases with Reverse-Transcriptase Activity in Coupled Reverse-Transcription Polymerase Chain Reaction Assays","abstract":"<jats:p>Reverse-transcription polymerase chain reaction (RT-PCR) is an important tool for the detection of target RNA molecules and the assay of RNA pathogens. Coupled RT-PCR is performed with an enzyme mixture containing a reverse transcriptase and a thermostable DNA polymerase. To date, several biotechnological companies offer artificial thermostable DNA polymerases with a built-in reverse-transcriptase activity for use in the coupled RT-PCR instead of the enzyme mix-tures. Here, we compared the artificial DNA polymerases and conventional enzyme mixtures for the RT-PCR by performing end-point and real-time RT-PCR assays using severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV2) RNA and endogenous mRNA molecules as tem-plates. We found that the artificial enzymes were suitable for different RT-PCR applications, in-cluding SARS-CoV2 RNA detection, but not for long-fragment RT-PCR amplification.</jats:p>","journal":null,"year":null,"id":653853,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1706121,"name":"Konstantin A. Blagodatskikh","orcid":null,"position":1,"is_corresponding":false},{"id":1706122,"name":"Ekaterina V. Barsova","orcid":null,"position":2,"is_corresponding":false},{"id":1706123,"name":"Dmitriy A. Varlamov","orcid":null,"position":3,"is_corresponding":false},{"id":1706124,"name":"Vladimir M. Kramarov","orcid":null,"position":4,"is_corresponding":false},{"id":1706125,"name":"Konstantin B. Ignatov","orcid":"0000-0001-8900-5278","position":5,"is_corresponding":false},{"id":1706120,"name":"Evgeniya V. Smirnova","orcid":"0000-0002-9744-952X","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Comparison of Commercially Available Thermostable DNA Polymerases with Reverse-Transcriptase Activity in Coupled Reverse-Transcription Polymerase Chain Reaction Assays","abstract":"<jats:p>Reverse-transcription polymerase chain reaction (RT-PCR) is an important tool for the detection of target RNA molecules and the assay of RNA pathogens. Coupled RT-PCR is performed with an enzyme mixture containing a reverse transcriptase and a thermostable DNA polymerase. To date, several biotechnological companies offer artificial thermostable DNA polymerases with a built-in reverse-transcriptase activity for use in the coupled RT-PCR instead of the enzyme mix-tures. Here, we compared the artificial DNA polymerases and conventional enzyme mixtures for the RT-PCR by performing end-point and real-time RT-PCR assays using severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV2) RNA and endogenous mRNA molecules as tem-plates. We found that the artificial enzymes were suitable for different RT-PCR applications, in-cluding SARS-CoV2 RNA detection, but not for long-fragment RT-PCR amplification.</jats:p>","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"19162232","pmcid":null,"openalex_id":"https://openalex.org/W4405715960","authors":[],"funders":[],"total_grants":0,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":"green","license":"cc-by","oa_locations":[{"url":"https://doi.org/10.20944/preprints202412.1876.v1","host_type":"repository"},{"url":"https://doi.org/10.20944/preprints202412.1876.v1","host_type":"repository"}],"fields_of_study":["SARS-CoV-2 detection and testing","Bacteriophages and microbial interactions","Viral gastroenteritis research and epidemiology"],"mesh_terms":[],"keywords":["Reverse transcriptase","Polymerase","Reverse transcription polymerase chain reaction","Molecular biology","RNA-Directed DNA Polymerase","RNA","DNA","Enzyme","Polymerase chain reaction","Biology","DNA polymerase","Real-time polymerase chain reaction","Inverse polymerase chain reaction","Messenger RNA","Chemistry","Biochemistry","Nested polymerase chain reaction","Gene"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-11T01:31:22.923750Z","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":[]}