Catharanthus roseus
Protocols: Cultures were washed and re-suspended as described (Schweizer et al., 2018). The suspensions were diluted to obtain a final concentration of OD600=0.4. Infiltration took place as described using four to five flowers, each from a different individual plant. Samples were harvested in four biological replicates as described (Schweizer et al., 2018). Opened flowers were removed two days prior to infiltration. A. tumefaciens pre-cultures were inoculated from glycerol stocks two days prior to infiltration to inoculate final overnight cultures one day prior to infiltration. About 100 mg of frozen ground plant material was used for RNA extraction using the Qiagen RNeasy mini kit. 450 μl Buffer RLT (supplemented with 10 μl of β-mercaptoethanol per ml Buffer RLT) was added and the mixture was vortexed vigorously. The lysate was transferred to a QIAshredder spin column placed in a 2 ml collection tube and centrifuged for 2 min at full speed. The supernatant was mixed with half a volume of ethanol and transferred to a an RNeasy spin column placed in a 2 ml collection tube followed by centrifugation for 15 seconds at full speed. 350 μl RW1 buffer was added and the column was centrifuged for 15s at max speed. Next an on column DNase digestion was performed and the column was washed again with 350 μl RW1 buffer. Next, the column was washed twice with 500 μl Buffer RPE. The RNA was eluated in a clean 1.5 mL eppendorf by adding 40 μl RNase-free water and centrifuging at maximum speed for 1 minute. Random primed cDNA library: Purification of poly-A containing mRNA molecules - mRNA fragmentation - Random primed cDNA synthesis - Adapter ligation and adapter specific PCR amplification.
ENA-FIRST-PUBLIC
2020-09-07T04:07:30Z
ENA-LAST-UPDATE
2020-09-04T10:40:08Z
External Id
SAMEA7290421
INSDC center name
Department of Plant Biotechnology and Bioinformatics, Ghent University, Gent, Belgium; VIB Center for Plant Systems Biology, VIB, Gent, Belgium
INSDC first public
2020-09-07T04:07:30Z
INSDC last update
2020-09-04T10:40:08Z
INSDC status
public
Submitter Id
E-MTAB-9517:S6_AGRO3
broker name
ArrayExpress
common name
Madagascar periwinkle
cultivar
little bright eyes
infect
Agrobacterium tumefaciens C58C1
organism part
petal
sample name
E-MTAB-9517:S6_AGRO3
scientific_name
Catharanthus roseus