Considering that Tempranillo berry skin and flesh shared circadian oscillatory gene expression (Additional file 2) and that a considerable portion of the ripening transcriptional program is common to both pericarp tissues [ 3], Tempranillo skin and flesh samples on each time point were considered as replicates to be compared to Verdejo pericarp samples.
Center fruit flesh samples were collected at stages of 10, 18, 28, and 34 DAP, respectively.
Soluble sugars were sampled using HPLC applying the procedure outlined at [ 62]; ethylene was sampled using GC-flame ionization detector (FID); pH of the flesh samples were measured by pH meter (PH-03 II), PH-03 IIument Corp., China) in juice squeeZD Instrument Corp.
Grape flesh samples (0.5 g) after certain procedures of preparation were used for determination of the soluble sugar content by HPLC (Agilent 1200 HPLC, USA), and the titratable acidity was determined by repeated titration with 0.1 mol/L NaOH (2 drops of 1% phenolphthalein added) to a faint pink.
Older laboratory techniques to identify fish meat looked at the mix of proteins in flesh samples, but were unreliable, expensive and cumbersome.
Two hundred milligrams of frozen watermelon flesh samples were ground to a fine powder prior to being extracted for 1 h in 10 ml of 50% ethanol at 80°C, then centrifuged at 3000 g for 10 min. This step was repeated one more time and the supernatants were then dissolved to a volumetric flask (25 ml) as extracts.
For all further analyses, flesh samples were taken from the heart area of each watermelon.
Approximately 10 ug cDNA from each of the four fruit flesh samples was used for sequencing on a Roche/454 GS-FLX Titanium instrument.
At the end of the trial (378 g average weight), flesh samples (Norwegian Quality Cut) were collected, frozen on dry ice and stored at −20°C until lipid analysis.
Flesh samples were taken by cutting two wedges, each approximately one-eighth of the fruit.
