2000
DOI: 10.1038/77367
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Enhanced detection and distinction of RNA by enzymatic probe ligation

Abstract: It is important that RNA molecules representing members of gene families are distinguished in expression analyses, and even greater resolving power may be required to identify allelic variants of transcripts in order to investigate imprinting or to study the distribution of mutant genes in tissues. Ligase-mediated gene detection allows precise distinction of DNA sequence variants, but it is not known if ligases can also be used to distinguish variants of RNA sequences. Here we present conditions for efficient … Show more

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Cited by 122 publications
(94 citation statements)
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“…T4 DNA ligase can join two DNA oligonucleotides using an RNA template (Kleppe et al 1970;Nilsson et al 2000). We therefore adopted an empirical approach to screen for Dand N-oligos for seven modification types.…”
Section: Finding D-and N-oligos For Seven Modification Typesmentioning
confidence: 99%
“…T4 DNA ligase can join two DNA oligonucleotides using an RNA template (Kleppe et al 1970;Nilsson et al 2000). We therefore adopted an empirical approach to screen for Dand N-oligos for seven modification types.…”
Section: Finding D-and N-oligos For Seven Modification Typesmentioning
confidence: 99%
“…[52][53][54][55] Although these methods are not yet available commercially, they have potentials to be developed into homogeneous and isothermal technologies that can overcome the barrier of PCR amplification.…”
Section: Thementioning
confidence: 99%
“…1; Nilsson et al 1994). Provided with the right conditions, DNA ligase will ligate the termini of the padlock probe on a perfectly matching RNA template, thereby accurately distinguishing matched and mismatched substrates (Nilsson et al 2000). The miRNA that is used as a template can subsequently be used as primer for rolling circle amplification, thereby linearly amplifying the target sequence in a process that has been shown to be highly quantitative ( Fig.…”
Section: Introductionmentioning
confidence: 99%