ArticlePloS one2013
Base preferences in non-templated nucleotide incorporation by MMLV-derived reverse transcriptases.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
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- The N-terminal region of malaria vaccine candidatebioRxiv : the preprint server for biology · 2025Article
- CRISPR with Transcriptional Readout reveals influenza transcription is modulated by NELF and can precipitate an interferon response.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- CRISPR with Transcriptional Readout reveals influenza transcription is modulated by NELF and can precipitate an interferon response.bioRxiv : the preprint server for biology · 2025Article
- Bacteroides sphingolipids promote anti-inflammatory responses through the mevalonate pathway.Cell host & microbe · 2025Article
- Analysis of NS2-dependent effects on influenza PB1 segment extends replication requirements beyond the canonical promoter.Nature communications · 2025Article
- Article
- Transcriptome and Temporal Transcriptome Analyses in Single Cells.International journal of molecular sciences · 2024Review
- Characterization and implementation of the MarathonRT template-switching reaction to expand the capabilities of RNA-seq.RNA (New York, N.Y.) · 2024Article
- Expression of thermostable MMLV reverse transcriptase in Escherichia coli by directed mutation.AMB Express · 2024Article
- Expression of Codon-Optimized Gene Encoding Murine Moloney Leukemia Virus Reverse Transcriptase in Escherichia coli.The protein journal · 2022Article
- Extensive age-dependent loss of antibody diversity in naturally short-lived turquoise killifish.eLife · 2022Article
- Rapid identification of CMV-specific TCRs via reverse TCR cloning system based on bulk TCR repertoire data.Frontiers in immunology · 2022Article
- A Molecular Analysis of Memory B Cell and Antibody Responses AgainstFrontiers in immunology · 2022Article
- Methyltransferase-directed orthogonal tagging and sequencing of miRNAs and bacterial small RNAs.BMC biology · 2021Article
- Persistence of Ambigrammatic Narnaviruses Requires Translation of the Reverse Open Reading Frame.Journal of virology · 2021Article
- Article
- Simple and efficient profiling of transcription initiation and transcript levels with STRIPE-seq.Genome research · 2020Article
- AP-TSS: A New Method for the Analysis of RNA Expression from Particular and Challenging Transcription Start Sites.Biomolecules · 2020Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Reverse transcriptases derived from Moloney Murine Leukemia Virus (MMLV) have an intrinsic terminal transferase activity, which causes the addition of a few non-templated nucleotides at the 3' end of cDNA, with a preference for cytosine. This mechanism can be exploited to make the reverse transcriptase switch template from the RNA molecule to a secondary oligonucleotide during first-strand cDNA synthesis, and thereby to introduce arbitrary barcode or adaptor sequences in the cDNA. Because the mechanism is relatively efficient and occurs in a single reaction, it has recently found use in several protocols for single-cell RNA sequencing. However, the base preference of the terminal transferase activity is not known in detail, which may lead to inefficiencies in template switching when starting from tiny amounts of mRNA. Here, we used fully degenerate oligos to determine the exact base preference at the template switching site up to a distance of ten nucleotides. We found a strong preference for guanosine at the first non-templated nucleotide, with a greatly reduced bias at progressively more distant positions. Based on this result, and a number of careful optimizations, we report conditions for efficient template switching for cDNA amplification from single cells.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.