Analysis of oligonucleotide microarrays by 3 ' end labeling using fluorescent nucleotides and terminal transferase

作者：Guerra Cesar E^*

来源：Biotechniques, 2006, 41(1): 53-56.

DOI：10.2144/000112182

A simple enzymatic labeling procedure is described to determine spot quality in oligonucleotide microarrays. By using fluorescently labeled dideoxynucleotides or ribonucleotides as substrate for terminal deoxynucleotidyl transferase (TdT), a single fluorophore can be covalently attached at the 3' end of each oligonucleotide probe molecule in the spot. Fluorescein-12-ddUTP Cy (TM) 3-ddUTP Cy5-UTP and Cy3-UTP were compared as TdT substrates for 3' end labeling an array of 1273 hexamer probes. Cy5-UTP was found to show minimal bias toward probe base composition and is therefore well suited for quantitative analysis of microarray spots where the oligonucleotide probes are coupled via a 5' end linkage to the solid phase.

• 出版日期2006-7