title: GSM6722776: F1R40 P3KO-11PEA-3; Mesocricetus auratus; ncRNA-Seq
library_construction_protocol: Samples were incubated at 72°C for 3 min to release and unfold the small RNAs or RNA. small RNA libraries construction: Briefly, Samples were incubated at 72°C for 3 min to release and unfold the small RNAs. After 3' adapter ligation, 5U of lambda exonuclease and 25U of 5' deadenylases, which performed better than the combination of RecJf and 5' deadenylases, were used to remove the excess 3' adaptor. Then, a 5' adapter was ligated and small RNAs were reverse transcribed. Pre-amplification was performed and then 1 μl of amplified product was used as a template for final amplification. The amplified libraries were separated on a 6% polyacrylamide gel and 130-160 bp DNA fragments were sliced to recover the library of small RNAs.  mRNA library construction:In brief, Samples were incubated at 72°C for 3 min to release the RNAs. For every single oocyte or embryo, 0.125 μL 1/100,000 dilution of the ERCC RNA Spike-In Mix (Invitrogen, Cat# 4456740) was used. Following reverse transcription and PCR pre-amplification, cDNA libraries were purified by Agencourt Ampure XP beads (0.8:1 ratio) (Beckman Coulter, Cat#A63881) and 2 ng of purified cDNA was used for a tagmentation reaction with Tn5 transposase. Amplified libraries were purified by Agencourt Ampure XP beads (1:1 ratio).