Nicotiana benthamiana
isolate
Laboratory strain
age
Leaves #3 and #4 at 2 days after agroinfiltration
geo_loc_name
Spain: Valencia
tissue
agroinfiltrated leaves
library_id
AC-HS-24
title
sRNA seq of N. benthamiana leaves agroinfiltrated with 35S:shc-amiR-NbSu
library_strategy
miRNA-Seq
library_source
TRANSCRIPTOMIC
library_selection
size fractionation
library_layout
single
platform
BGISEQ
instrument_model
DNBSEQ-G400
design_description
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
filetype
fastq
filename
AC-HS-24_1.fq.gz
BioSampleModel
Plant