Terminal Deoxynucleotidyl Transferase Mediated Production of Labeled Probes for Single-molecule FISH or RNA Capture

被引:17
作者
Gaspar, Imre [1 ]
Wippich, Frank [1 ]
Ephrussi, Anne [1 ]
机构
[1] European Mol Biol Lab, Dev Biol Unit, Meyerhofstr 1, D-69117 Heidelberg, Germany
关键词
Terminal deoxynucleotidyl transferase; Labelled terminator nucleotide; Probe production; Single molecule FISH (smFISH); RNA capture; RNA affinity purification (RAP);
D O I
10.21769/BioProtoc.2750
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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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页数:20
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