Sample 32
Euphorbia pulcherrima
Protocols: Cuttings with two small leaves were taken from Euphorbia pulcherrima Willd ex Klotzsch. (Poinsettia) 'Lilo Red' (Ecke Ranch, California, USA) mother plants grown under long day conditions (LD) (16 h light from High Pressure Sodium (HPS lamps) at 150 molm-2s-1, 22°C day/ 20°C night, 70% Relative Humidity (RH)).The freeze-dried material on the slide PET membranes were placed on the laser microdissection microscope (Leica model 6000, Microsystems, Germany), the objective 6,3X was used to dissect the three different tissues for further RNA isolation: distal, abscission zone (AZ) and proximal. The software Laser microdissection software V6.7.2.4295 (Leica Microsystems, Germany), was used to control the microscope and select the average conditions of the laser such as the aperture 45, laser power 60 and speed 20. The mass of the tissues was calculated using the volume obtained. The microdissected tissues were collected inside a cap of a 0,6 ml RNAse free microcentrifuge tube, where they fell by gravity. The RNA isolation from the microdissected tissues was performed using the Ambion RNAqueous®- Micro kit (Ambion Inc., Austin, TX,Country??) following supplier instructions with small modifications as the use of a tip to crash the material inside the tube in presence of the lysis buffer. The protocol of isolation was followed by a 20 minutes DNAse treatment according to supplier instructions. The RNA was resuspended in 20  of nucleases free water, purified by LiCl-EtOH precipitation and stored at -80°C. The concentration and quality of the RNA was analysed using the Agilent 2100 Bioanalyzer (company, country) and the RNA 6000 Pico Kit chip (Agilent Technologies, CA, USA), following manufacturer instructions. The libraries were prepared according to the TruSeq RNA Sample Prep Kit v2 (Illumina, San Diego, CA, USA)
ENA first public
2017-12-31
ENA last update
2017-12-06
External Id
SAMEA104417725
INSDC center alias
Department of Plant Sciences, Faculty of Biosciences, Norwegian University of Life Sciences
INSDC center name
Department of Plant Sciences, Faculty of Biosciences, Norwegian University of Life Sciences
INSDC first public
2017-12-31T17:01:47Z
INSDC last update
2017-12-06T12:11:53Z
INSDC status
public
Submitter Id
E-MTAB-6282:Sample 32
broker name
ArrayExpress
common name
poinsettia
cultivar
Lilo Red
growth condition
long day length regimen
organism part
flower bud
sample name
E-MTAB-6282:Sample 32
sampling site
proximal flower bud