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Grains from 0, 4, 8, 10, 12, 14 DAF were manually dissected into maternal and filial parts and whole caryopses were sampled at 16, 20 and 24 DAF.
In brief, 2 mm skin punch biopsies were manually dissected into five to ten tissue pieces and transferred to 1.5 ml medium in a 25-cm cell culture flask (BD Falcon, Heidelberg, Germany).
Skin punch biopsies (3 mm) were rinsed in DPBS containing Primocin (InvivoGen, San Diego, California) and penicillin/streptomycin (Pen/Strep, Corning, Corning, New York) and manually dissected into 10 15 smaller pieces.
For seed compartments, fresh seeds were manually dissected into radical, hypocotyl, cotyledon, embryo, endosperm and seed coat in RNAlater (Life Technologies) and kept at 4°C for a few days for the solution to penetrate into the seeds and then stored in −20°C.
Whole UCs were manually dissected into smaller sections (5-6 grams UC tissue per isolation) and plated in polystyrene tissue culture flasks with L-DMEM supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin/streptomycin (Gibco, Carlsbad, CA, USA) for 7 days in a 37°C humidified environment with 5% CO2 [ 24].
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For library construction, all nectaries were manually dissected from B. rapa flowers (an example dissection is available in Video S1), and processed as described in Materials and Methods.
The tumor was manually dissected from each unstained slide using a scalpel blade and transferred into a microcentrifuge tube.
The fruit AZs, located between the pedicel and fruit, were manually dissected from longitudinal sections of the samples with a razor blade into pieces to a maximum width of 1 mm on each side of the abscission fracture plane [ 15].
A rat kidney is perfused with and incubated in collagenase solution, and the digested kidney was manually dissected under a stereomicroscope.
To validate the bioinformatics tools, we randomly sampled a subset of the total data and manually dissected the raw reads one by one to identify the CDR3 sequences.
The maternal basal plate (decidua) was manually dissected from the fetal placental tissue.
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