@article {Gaspar107599, author = {Imre Gaspar and Frank Wippich and Anne Ephrussi}, title = {Enzymatic production of single molecule FISH and RNA capture probes}, elocation-id = {107599}, year = {2017}, doi = {10.1101/107599}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Arrays of singly-labelled short oligonucleotides that hybridize to a specific target revolutionized RNA biology, enabling quantitative, single molecule microscopy analysis and high efficiency RNA/RNP capture. Here, we describe a simple and efficient method that allows flexible functionalization of inexpensive DNA oligonucleotides by different fluorescent dyes or biotin using terminal deoxynucleotidyl transferase and custom-made functional group conjugated dideoxy-UTP. We show that 1) all steps of the oligonucleotide labelling {\textendash} including conjugation, enzymatic synthesis and product purification {\textendash} can be performed in a standard biology laboratory, 2) the process yields \>90 \%, often \>95 \% labeled product with minimal carry-over of impurities and 3) the oligonucleotides can be labeled with different dyes or biotin, allowing single molecule FISH or RNA affinity purification to be performed.}, URL = {https://www.biorxiv.org/content/early/2017/02/10/107599}, eprint = {https://www.biorxiv.org/content/early/2017/02/10/107599.full.pdf}, journal = {bioRxiv} }