GSM5229041: RppH_minus (Intact); Mus musculus; ncRNA-Seq
Total brain RNA was prepared by using isogene. Five hundred nanograms of RNAs were treated (or not treated as a control) with RNA 5' Pyrophosphohydrolase (RppH, New England Biolab) for 1 hour at 37 ºC in a 50-μl reaction mixture containing 1X Thermopol buffer (New England Biolab), and the reaction was stopped by phenol-chloroform extraction and ethanol precipitation. NEBNExt small RNA library preparation kit (New England Biolab) was then used for constructing sequence libraries.  The libraries were run on a p% polyacrylamide gel and DNAs of 240 to 380 bp (corresponding to insert size of 113 to 253 bp) were purified from the gel. The libraries were sequenced on MiSeq with MiSeq Reagent Kit v2 (300 cycles) in the 300-bp single-end mode