Chemical Modification of Oligonucleotides: A Novel Approach Towards Gene Targeting

Torigoe, Hidetaka; Imanishi, Takeshi

doi:10.5772/50393

Mutagenesis 2012

DOI: 10.5772/50393

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Chemical Modification of Oligonucleotides: A Novel Approach Towards Gene Targeting

Hidetaka Torigoe¹,

Takeshi Imanishi²

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2016

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“…The universal priming ONT is also 5-phosphorylated and contains a sequence complimentary to the non-telomeric sequence in the template ONT with additional thymine and adenine bridged nucleic acids (BNAs) at the 3 end (Supplementary Table S1) such that it anneals to the template ONT in the correct spot. We decided to use BNA-containing ONTs since they have a greater resistance to nuclease digestion and a higher affinity for the target DNA or RNA as compared with locked nucleic acids (9,10). …”

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Generation of Digoxigenin-Incorporated Probes to Enhance DNA Detection Sensitivity

Lai

Wright

2016

BioTechniques

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Telomere length in humans has been correlated with cancer and age-related diseases. The standard method to measure telomere length relies on Southern blot analysis with radioactively or non-radioactively labeled probes containing several telomeric DNA repeats. However, this approach requires relatively large amounts of genomic DNA, making it difficult to measure telomere length when a limited amount of sample is available. Here, we describe a non-radioactive labeling method that uses 3' fill-in combined with lambda exonuclease digestion to incorporate one or more digoxigenin (DIG) molecules into bridged nucleic acid (BNA)-containing oligonucleotides (ONTs). Using our method, we were able to generate probes to detect both C- and G-rich telomeric DNA strands. Compared with commercially available DIG-labeled telomere probes, probes generated using this new approach significantly enhance the sensitivity of telomere length measurements.

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mentioning

confidence: 99%

Generation of Digoxigenin-Incorporated Probes to Enhance DNA Detection Sensitivity

Lai

Wright

2016

BioTechniques

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Chemical Modification of Oligonucleotides: A Novel Approach Towards Gene Targeting

Cited by 1 publication

References 41 publications

Generation of Digoxigenin-Incorporated Probes to Enhance DNA Detection Sensitivity

Generation of Digoxigenin-Incorporated Probes to Enhance DNA Detection Sensitivity

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