Efficient microRNA capture and bar-coding via enzymatic oligonucleotide adenylation.
Here we report a highly efficient and simplified strategy to preadenylate bar-coded oligonucleotides designed for microRNA (miRNA) capture and multiplex analysis. Using this approach, we enzymatically preadenylated bar-coded oligonucleotides with high efficiency when compared to the chemical method currently used by miRNA investigators. As a case study, we used these oligonucleotides in an ATP-independent ligation to miRNAs, suggesting the utility of our method in end-capture protocols and high-throughput sequencing applications.
Vigneault
Francois
F
Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA. fvigno@hms.harvard.edu
Sismour
A Michael
AM
Church
George M
GM
D000225:Adenine / Q000737:chemistry*; D011088:DNA Ligases / Q000378:metabolism*; D001330:Electronic Data Processing; D006801:Humans; D035683:MicroRNAs / Q000032:analysis*; D009841:Oligonucleotides / Q000737:chemistry*; D011130:RNA Ligase (ATP) / Q000378:metabolism*
D035683:MicroRNAs; D009841:Oligonucleotides; D011088:DNA Ligases; D011130:RNA Ligase (ATP); D000225:Adenine