Generation of cDNA libraries from RNP-derived regulatory noncoding RNAs.
Next-generation sequencing of noncoding RNA (ncRNA) libraries has become an essential tool for the profiling of ncRNAs and the identification of novel ncRNA species. Here, we describe the generation of a ncRNA-derived complementary DNA (cDNA) library by 3'-tailing of ncRNAs by CTP and poly(A) polymerase, followed by 5'-adapter ligation by T4 RNA ligase and reverse transcription of ncRNAs with an oligo-d(G) anchor primer. Preliminary selection of ncRNAs from ribonucleoprotein particles (RNPs) enables a strong enrichment of the generated libraries with functional regulatory ncRNAs compared to classical approaches.
Biopôle, CNRS UMR 7214 AREMS, Université de Lorraine, Vandoeuvre-lès-Nancy, France. mathieu.rederstorff@maem.uhp-nancy.fr
Mathieu
M
Rederstorff
D003570:Cytidine Triphosphate / Q000378:metabolism; D015723:Gene Library; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000302:isolation & purification / Q000378:metabolism*
D022661:RNA, Untranslated; D012261:Ribonucleoproteins; D003570:Cytidine Triphosphate; D011062:Polynucleotide Adenylyltransferase