title: GSM4305663: At ddm1 pol V; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: Total RNA was extracted from frozen inflorescence tissue using TriZOL LS protocol (Invitrogen) and enriched for poly(A) mRNA using NEBNext Poly(A) mRNA Magnetic Isolation Module. SQK-PCS108 protocol by Oxford Nanopore was used to construct the library. 50 ng of enriched poly(A) mRNA was used for reverse transcription. 14 cycles of PCR amplification was performed with barcoding adapters as per SQK-PBK004 protocol by Oxford Nanopore. 1D sequencing adapter was added before sequencing on a SpotON flow cell in MinION sequencer. For the higher ddm1/rdr6/pol V rep B, only 12 cycles of amplification was used. Multiplxed library of 5 genotypes was sequenced in Round 1. For round 2 with higher depth ddm1/rdr6/pol V sample, only 1 barcode was used.
attributes.tag: GEO Accession
attributes.value: GSM4305663