LIBEST_028273
External Id
SAMN02340059
External Id
National Research centre On Plant Biotechnology
INSDC center name
2014-06-30T00:44:09.183Z
INSDC first public
2017-10-11T17:48:31.340Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
BioYSR
cultivar
Plants of Indian mustard were raised under growth chamber conditions at National Phytotron Facility, Indian Agricultural research Institute, New Delhi, India. About 21 days old plants were sprayed with 50 mM Methyl jasmonate (MeJ) in 0.001% triton X100 and along with control plants homologously treated with 0.001% triton X100 in water. Leaves were harvested and flash frozen in liquid nitrogen at 0h and 1h after the treatments and stored at -80C until further used. Total RNA was isolated from the frozen samples with Trizol reagent (Invitrogen) and mRNA was fractionated from total RNA by using Magnetic mRNA Isolation Kit (New England Biolabs, UK) according to manufacturer's protocol. Subtracted cDNA library was constructed by using PCR-Select Subtractive Hybridization kit (DSS Takara Bio India Pvt. Ltd.). cDNA prepared from the MeJ treated sample was used as the 'tester' and water treated control sample as 'driver' for the construction of forward subtraction library. The SSH cDNA library was generated by ligating subtracted cDNAs into pCR2.1 topo T/A cloning vector. From the library differentially expressed clones were identified by macro array analysis and selected clones were sequenced by using universal M13 forward primer. Sequence data were processed with DNA star software to remove vector and adaptor sequence contamination to obtain high quality reads. The sequences were deposited to genbank database.
description
21 days old plants
dev-stage
E.coli DH5 alpha
lab-host
leaf
http://purl.obolibrary.org/obo/PO_0025034
organ
Brassica juncea
http://purl.obolibrary.org/obo/NCBITaxon_3707
organism
bisexual
sex
leaf
http://purl.obolibrary.org/obo/PO_0025034
tissue-type
SSH cDNA library of methyl jasmonate treated Indian mustard
title
Plasmid
http://purl.obolibrary.org/obo/SO_0000155
v_type
pCR 2.1 Topo T/A cloning vector
vector