Based on microscopy inferences, 48 HAP pistils of SP_S, SP_T and CP (SxT) in ten biological replicates were collected and snap-frozen to liquid nitrogen for total RNA extraction using IRIS method [51 (link)]. The RNA was quantified on NanoDrop 2000 (Thermo Scientific, Waltham, MA, USA), and quality was assessed on 1% formaldehyde agarose gel (MOPS) and Agilent Bioanalyzer with RNA 7500 series II Chip (Agilent Technologies, CA, USA). The RNA samples with RIN (RNA Integrity Number) value greater than 8 and the final concentration of 4.0 µg were used for cDNA library preparation.
Eight cDNA libraries in biological replicates SP_T (3), CP (3) and SP_S (2) were constructed using the illumina Truseq RNA Sample prep v2 LS Protocol (Illumina Inc., CA, USA). The libraries were quantified on Qubit 2.0 fluorometer (Invitrogen, USA), while quality was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA). The paired-end (PE) (2 × 72 bp) sequencing was performed using Illumina GAIIx.
Eight cDNA libraries in biological replicates SP_T (3), CP (3) and SP_S (2) were constructed using the illumina Truseq RNA Sample prep v2 LS Protocol (Illumina Inc., CA, USA). The libraries were quantified on Qubit 2.0 fluorometer (Invitrogen, USA), while quality was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA). The paired-end (PE) (2 × 72 bp) sequencing was performed using Illumina GAIIx.
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