title: CRAC of Xrn1 Tail of S. cerevisiae
design: Crosslinking and Analysis of cDNA (CRAC): BY4741 yeast strains expressing His6-TEV protease cleavage site- ProteinA (HTP) tagged Xrn1 or Xrn1 tail were grown in exponential phase before crosslinking using UV light at 254 nm. Cells were lysed and RNA-protein complexes were first retrieved on IgG sepharose and then on NiNTA in the presence of 6 M guanidium-HCl. RNAs were trimmed using RNace-IT (see Bohnsack et al., 2012). Illumina sequencing adaptors (RA3 TGGAATTCTCGGGTGCCAAGG-3) and RA5 (5-GTTCAGAGTTCTACAGTCCGACGATCNNNNNAGC-3) were ligated. cDNA libraries were prepared by reverse transcription using Superscript III and amplified by PCR. Following agarose gel electrophoresis, libraries were size selected (150-300 nt) and purified.