GSM4728977: 5Pase; Mus musculus; RNA-Seq
Total testes RNA was prepared by using isogene. Six hundred nanograms of RNAs were treated (or not treated as a control) with 25 U of tobacco alkaline phosphatase (TAP) (Epigcentre) for 1.5 hours at 37 ºC, and the reaction was stopped by phenol-chloroform extraction and ethanol precipitation. The total intact RNA was also treated with 20 U of 5' polyphosphatase (5Pase) (Epicentre) for 30 min. at 37 ºC, and the reaction was stopped by phenol-chloroform extraction and ethanol precipitation. TruSeq small RNA library preparation kit (Illumina) was then used for constructing sequence libraries. The libraries were run on a 2% agarose gel and DNAs of 240 to 380 bp (corresponding to insert size of 115 to 255 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.