SAMD00189826
External Id
Laboratory of Gene Expression, Medical Research Institute, Tokyo Medical and Dental University
INSDC center name
2021-01-29T00:00:00Z
INSDC first public
2021-01-29T08:17:11Z
INSDC last update
DRS171743
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
DRS171743
SRA accession
sams-3 and sams-5 genomic DNA fragments were amplified from N2 genomic DNA with primer pairs 5-GTTTCAACCTTACAACCAAA-3 and 5-CGGACGAGAACYTGRTAGTC-3, and 5-ACAAAGCTTCCCCAGCAAGT-3 and 5-CGGACGAGAACYTGRTAGTC-3, respectively, by using PrimeStar GXL (Takara). The sams-3 and sams-5 DNA fragments were further amplified with a forward primer 5-TAATACGACTCACTATAGGGAAACGGGGCTGAAGC-3 (sams-3) or 5-TAATACGACTCACTATAGGGTGACAAGAGGATGAAAC-3 (sams-5) and a reverse primer 5-TTTTTTTTTTTTTTTTTTTTGGGGGGGGGATGACGTTTCGGACGAGAAC-3 (C6A20) or 5-TTTTTTTTTTTTTTTTTTTTCCCCCCCCCATGACGTTTCGGACGAGAAC-3 (G6A20) to attach a T7 promoter and a C9A20 or G9A20 tail. The four DNA fragments were utilized as templates for in vitro transcription with T7 RNA polymerase and the RNAs were purified with PAGE and gel extraction. The G9A20-tagged sams-3 and sams-5 RNAs were subjected to methylation reaction with recombinant human METTL16 MTD. The four RNAs were purified again with PAGE and gel extraction. The four RNA samples were then pooled and subjected direct RNA sequencing with Nanopore MinION platform.
description
wild-type
http://www.ebi.ac.uk/efo/EFO_0005168
genotype
Caenorhabditis elegans
http://purl.obolibrary.org/obo/NCBITaxon_6239
organism
WT
http://www.ebi.ac.uk/efo/EFO_0001103
phenotype
sams-345
sample name
N2
http://www.ebi.ac.uk/efo/EFO_0004028
strain
in vitro transcribed RNA
tissue
C.elegans_N2_genome_sams-3,5_in-vitro-transcription_METTL16
title