Terminal deoxynucleotidyl transferase mediated production of labeled probes for single-molecule FISH or RNA capture

Authors:

Gaspar, I., Wippich, F., & Ephrussi, A.

Keywords:

Terminal deoxynucleotidyl transferase, Labelled terminator nucleotide, Probe production, Single molecule FISH (smFISH), RNA capture, RNA affinity purification (RAP)

Abstract:

Arrays of short, singly-labeled ssDNA oligonucleotides enable in situ hybridization with single molecule sensitivity and efficient transcript specific RNA capture. Here, we describe a simple, enzymatic protocol that can be carried out using basic laboratory equipment to convert arrays of PCR oligos into smFISH and RAP probesets in a quantitative, cost-efficient and flexible way.

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