LIBEST_028283
External Id
SAMN02364163
External Id
Tea Research Institute Chinese Academy of Agricultural Sciences (TRICAAS)
INSDC center name
2013-09-25T23:05:03.713Z
INSDC first public
2015-05-20T15:03:32.377Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
Longjing 43
cultivar
Freshly dormant axillary buds and sprouting buds of Camellia sinensis var. Longjing 43, a special early sprouting cultivar, were collected from the tea garden of Tea Research Institute Chinese Academy of Agricultural Sciences, Hangzhou, China. Total RNA was extracted from both samples and ds cDNA was synthesised using the SMART cDNA library construction kit (Clontech Laboratories Inc., CA, USA). The PCR based cDNA subtraction method, suppression subtractive hybridization (SSH) was performed with both samples using the PCR-Select cDNA subtraction kit (Clontech,USA). The cDNA fragments generated from the mRNA of both dormant buds and sprouting buds were used as tester and driver respectively. The SSH library enriched for differentially expressed cDNA was constructed by ligation of the subtracted cDNAs into the pMD18-T vector (Takara, Dalian).The ligation mixture was transformed into competent E.coli cells and plated onto LB agar media and incubated overnight to obtain a subtracted EST bank. Plasmid DNA was extracted from randomly selected white EST colonies and grown in liquid LB media for 24 hours. DNA sequencing was performed using an automatic DNA sequencer (ABI prism).
description
dormant buds
dev-stage
E. coli
lab-host
Camellia sinensis
http://purl.obolibrary.org/obo/NCBITaxon_4442
organism
buds
http://purl.obolibrary.org/obo/PO_0000055
tissue-type
LIB-EST LJDB (Longjing 43 dormant buds) Camellia sinensis subtractive cDNA library
title
pMD18-T vector
vector