GSM8785618: Col-0 meiocyte replicate 5; Arabidopsis thaliana; RNA-Seq
Anthers of col-0, c3h14, c3h15 and c3h14 c3h15, during anther stage 4 to 7, were collected respectively and squeezed with forceps to release meiocytes. Recombinant RNase inhibitor (final concentration with 1 U/μL, Takara, 2313B) was used to prevent RNA degradation by adding to the collection buffer. Then RNA of meiocytes was extracted by TRIzol reagent (ThermoFisher, 15596026), facilitated by Dr. GenTLE™ Precipitation Carrier (Takara, 9094) for better precipitation of microscale RNA. As for inflorescence meristem (IM) and anthers at stage 2 from col-0, tissues were isolated and collected from Arabidopsis inflorescences. Leaves from col-0 and c3h14 c3h15 that had grown for 21 days were also collected for RNA-seq. All these tissue samples were used to extract RNA by using the TRIzol reagent (ThermoFisher, 15596026). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.