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Sample GSM1855472 Query DataSets for GSM1855472
Status Public on Feb 10, 2017
Title Xb leaves
Sample type SRA
 
Source name boron-sufficient leaves
Organism Citrus sinensis
Characteristics tissue: Fully expanded Citrus sinensis leaves were excised from the seedlings
treatment: boron-sufficient
Treatment protocol Xa(B-deficient ,0 μM H3BO3),Xb(B-sufficient ,10 μM H3BO3)
Growth protocol 5-week-old seedlings of ‘Xuegan’ [Citrus sinensis (L.) Osbeck] were transplanted into 6 L pots containing fine river sand. Ten weeks after transplanting, each pot was supplied every other day with B-deficient (0 μM H3BO3) or B-sufficient (10 μM H3BO3, control) nutrient solution for 15 weeks. There were 10 replications per B treatment with 2 pots in a completely randomized design.
Extracted molecule total RNA
Extraction protocol About 0.1 g mixed frozen B-sufficient and B-deficient leaves from five replictations were used to extract RNA. Total RNA was extracted from frozen roots using TRIzol reagent (Invitrogen, Carlsbad, CA)
Small RNA samples were sequenced by Beijing Genomics Institute (BGI) (Shenzhen, Guangdong, China) using the high throughput pyrosequencing technology developed by Illumina. Two sRNA libraries were constructed, sRNAs were isolated from the total RNA by size fractionation with 15% Tris-borate-EDTA urea polyacrylamide gel (TBU). Then the sRNAs were ligated with 5' and 3' adaptor by T4 RNA ligase after being dephosphorylatd by alkaline phosphatase. The adaptor-ligated sRNAs were transcribed to single-stranded cDNA using Superscript II reverse transcriptase (Invitrogen). Thereafter, the single-stranded cDNA was used as templates for double-stranded synthesis by PCR amplification using the primer designed according to the adapter sequence. The obtained PCR products were sequenced on a Solexa sequencer (Illumina) at the Beijing Genomics Institute (BGI), Shenzhen, China.
 
Library strategy miRNA-Seq
Library source transcriptomic
Library selection size fractionation
Instrument model Illumina Genome Analyzer
 
Data processing Basecalls performed
Get rid of some contaminant reads from the fq file and get the final clean reads.
The clear reads were mapped to Citrus clementina genome (JGI version 1.1, http://phytozome.jgi.doe.gov/pz/portal.html#!info?alias=Org_Csinensis) using SOAP, only perfectly mapped sequences were retained and analyzed further. rRNAs, tRNAs, snRNAs and snoRNAs were removed from the sRNAs sequences through BLASTn search using NCBI Genebank database (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi/) and Rfam (12.0) database (http://www.sanger.ac.uk/resources/databases/rfam.html) (e = 0.01). The remaining sequences were aligned with known plant miRNAs from miRBase 21 (http://www.mirbase.org/).
Reads that were not annotated were used to predict novel miRNAs using a prediction software Mireap (http://sourceforge.net/projects/mireap/), which was developed by the BGI
Supplementary_files_format_and_content: abundance
 
Submission date Aug 16, 2015
Last update date May 15, 2019
Contact name Chen Lisong
E-mail(s) [email protected]
Phone +8613599390418
Organization name Fujian Agriculture and Forestry University
Street address Shangdian street NO.15
City Fuzhou
ZIP/Postal code 350002
Country China
 
Platform ID GPL9256
Series (1)
GSE72108 Boron-deficiency-responsive microRNAs and their targets in Citrus sinensis leaves
Relations
BioSample SAMN03995873
SRA SRX1153148

Supplementary file Size Download File type/resource
GSM1855472_Xb_clean.txt.gz 29.8 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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