| Growth Protocol: |
Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~1 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~2 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~3 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~4 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~5 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~6 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~7 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~8 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~9 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~10 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~11 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~12 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~13 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~14 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers.; Arabidopsis plants were grown in a growth chamber under the following conditions: long-day photoperiod (16-h light/8-h dark), light intensity 170 μmoles m-2 s-1, day/night temperature 21°C/17 °C. Shortly after bolting (with stem length ~15 cm), the shoot apices were cut and the SAMs were dissected by removing fully open flowers. |
| Extract Protocol: |
Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2200 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2201 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2202 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2203 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2204 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2205 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2206 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2207 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2208 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2209 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2210 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2211 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2212 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2213 TapeStation.; Total RNA was isolated from 36 dissected inflorescence meristems using RNeasy Mini Kit (QIAGEN Cat No./ID: 74104), following the manufacturer’s instructions. RNA quality and integrity was assessed on the Agilent 2214 TapeStation. |
| Library Construction Protocol: |
After the QC procedures, mRNA from eukaryotic organisms is enriched using oligo(dT) beads. For prokaryotic organisms or eukaryotic organisms' long-non-coding libraries, rRNA is removed using the Ribo-Zero kit that leaves the mRNA. First, the mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.The qualified libraries are fed into Illumina sequencers after pooling according to its effective concentration and expected data volume. |
