P-MTAB-32855 - P-MTAB-32855
Rapid Amplification of cDNA Ends
5' and 3' RACE reactions were performed in 12.5 ul volume independently with each polyA+ RNA sample using the SMART RACE amplification kit (Clontech) according to the manufacturer’s instructions. 7.5ul of each of the 400 lncRNA targets were pooled respecting tissues and directionality of the RACE reaction (5' and 3' RACE) and purified with the QIAquick kit (Quiagene) as described for RT-PCR-seq experiments . A set of 25 control reactions targeting full-length annotated transcripts were RACEd in parallel as positive controls. 1.5ul of these controls were pooled with the other reactions prior to library preparation.