GSM2869263: Mtr4_CRAC_rep2; Mus musculus; OTHER
2x15cm dishes of mESCs were crosslinked, cells lysed in mild RIPA buffer, a tandem purification with FLAG and then Streptavidin beads performed, including RNase digestion, TSAP treatment, PNK treatment, and linker ligations while on the Streptavidin beads. The RNPs were eluted by heating in loading buffer, and resolved by SDS-PAGE and transfer to a nitrocellulose membrane. The region corresponding to the (radiolabelled) RNP was then cut out, RNA extracted using Proteinase K and then phenol:choloroform, and then cDNA synthesis and PCR performed. The final library was purified on a Metaphor agarose 3 % gel. The 5' linker includes an in-line barcode, followed by AGC at the 3' end.
GEO Accession
GSM2869263