GSM5068444: AZ_3_150; Elaeis guineensis; RNA-Seq
For RNA extractions, the fruit bunches were collected and the base of fruits containing the AZ were sampled as follows: the spikelets were removed from the bunches, rinsed in water, then individual fruit containing the base of the fruit containing the AZ were removed with scalpel, then the base of the fruit containing the AZ were dissected (approximately 50 AZ pieces were collected per sample), weighed at approximately 3 grams then froze immediately in liquid nitrogen. Backcross material were cleaned and processed with the same procedure. Total AZ weights were approximately 3-6 grams. Total RNA from mesocarp, pedicel and the base of the fruit enriched in AZs, treated or not with ethylene was extracted as previously described [Morcillo F et al.,Tree Physiol. 2006;26(5):585-594]. Total RNA (1 μg) was used to synthesize cDNA using the first-strand cDNA synthesis kit (ImProm-IITM Reverse Transcription System, Promega). The titanium kit was used (according to Roche GS FLX Titanium manual) and cDNAs derived from the different tissues and different ethylene treatment time points were tagged independently and then mixed together in one sample for 454 pyrosequencing carried out by the National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand. Sequencing of the three natural abscission samples (AZ30, AZ120 and AZ160) collected in Thailand was complete by the company GATC Biotech (www.gatc-biotech.com).