GSM5856599: KO-33PEA smRNA rep1; Mesocricetus auratus; ncRNA-Seq
The single oocytes or embryos were incubated at 72°C for 3 min to release and unfold the small RNAs.For testicular RNA seq, total RNA was extracted from 21 d.p.p. golden hasmters using the RNA isolater Total RNA Extraction Reagent(VAZYME,R401-01)according to the manufacturer's protocol. For single oocyte or embryos smRNA seq, 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. Long RNA transcriptome libraries was performed following Smarter-seq2 method with some modifications.For every single oocyte or embryo, 0.125 μ