LIBEST_028187
External Id
SAMN02086934
External Id
College of Horticulture and Forestry
INSDC center name
2014-04-23T00:44:08.933Z
INSDC first public
2017-10-11T17:48:29.500Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
Fantasy Red
cultivar
In March of 2010, seeds of Petunia hybrida 'Fantasy Red' were sown into plug-trays. When the crop canopy closed, single plants were transferred to 9 cm pots. The plants were treated under short days in a growth chamber at 23 C day /18 C night and 60 +/- 10% relative humidity until abnormal flower buds appeared. To prepare a representative sample of total RNA from the normal and abortion flower buds in SSH process, roughly the same size of plant organs (3 mm +/- 0.5) were pooled, respectively. SSH library was performed using PCR Select cDNA Subtraction kit (Clontech, USA) according to the manufacturer's instructions. The final PCR products were purified using AxyPrepTM DNA Gel Extraction kit (Axygen Scientific, USA). The fragments obtained were cloned into the pMD18-T vector (TaKaRa, Japan) and used for the transformation of Escherichia coli DH5alpha (TransGen Biotech, Beijing). A total of 4032 and 3840 clones were randomly selected to construct the forward and reverse SSH libraries, respectively. One microliter of each bacterial culture was used for the PCR amplification with nested PCR primers 1 (5'-TCGAGCGGCCGCCCGGGCAGGT-3') and 2R (5'-AGCGTGGTCGCGGCCGAGGT-3'). The PCR program is: 95 C for 5 min; 94 C for 30 s, 61 C for 40 s, 72 C for 1 min, 30 cycles followed by a 5 min final extension at 72 C.
description
Petunia x hybrida
http://purl.obolibrary.org/obo/NCBITaxon_4102
organism
flower bud
http://purl.obolibrary.org/obo/PO_0000056
tissue-type
The forward subtracted cDNA library of short-day induced petunia abortion flower buds
title