35S.pri-amiR-NbSu (agroinfiltrated leaves)
SAMN34247085
External Id
Instituto de Biologia Molecular y Celular de Plantas
INSDC center name
2023-12-18T00:00:00Z
INSDC first public
2023-12-18T08:11:02.361Z
INSDC last update
SRS17351374
INSDC secondary accession
live
INSDC status
Plant
NCBI submission model
Plant.1.0
NCBI submission package
SRS17351374
SRA accession
Leaves #3 and #4 at 2 days after agroinfiltration
age
Steps of library preparation by BGI:1) Small RNA selection: Use PAGE electrophoresis to cut and separate RNA of size 18-30nt from total RNA;2) 3' adapter ligation: Use 5-adenylated and 3-blocked single stranded DNA adapter to ligate with the 3' end of RNA fromstep (1);3) Annealing of reverse transcription primers: Add RT primers into the system from step (2), hybridize with the 3'adapter attached to the RNA, meanwhile hybridize with excess 3' adapter;4) 5' adapter ligation: Ligate the 5' adapter to the 5' end of products from step (3). Since the adapter preferentially ligateswith single-stranded molecules rather than the hybrid of 3' adapter and RT primer, it can greatly reduce the ligationbetween adapters;5) Synthesize the first strand of cDNA: Use RT primer from step (3) for reverse transcription extension, thensynthesize the first strand of cDNA;6) PCR amplification: Use high-sensitivity polymerase to amplify cDNA, enrich the cDNA with both 3' adapter and 5'adapter, then amplify the library yield;7) Library fragment selection: Use PAGE electrophoresis to separate PCR products in the range of 100-120bp, whicheffectively removes by-products such as primer dimers;8) Library quantification and pooling cyclization
design_description
AC-HS-22_1.fq.gz
filename
fastq
filetype
Spain: Valencia
geo_loc_name
DNBSEQ-G400
instrument_model
Laboratory strain
isolate
AC-HS-22
library_id
single
library_layout
size fractionation
library_selection
TRANSCRIPTOMIC
library_source
miRNA-Seq
library_strategy
Nicotiana benthamiana
http://purl.obolibrary.org/obo/NCBITaxon_4100
organism
BGISEQ
platform
agroinfiltrated leaves
tissue
Plant sample from Nicotiana benthamiana
title
sRNA seq of N. benthamiana leaves agroinfiltrated with 35S:pri-amiR-NbSu
title