ena-STUDY-CHINA Wuhan University-24-01-2018-10:00:55:438-46
This study presented the transcriptome sequencing of submerged and floating leaves of the aquatic plant P. octandrus.
P. octandrus total RNA was extracted from submerged and floating leaves. 10 µg of total RNA was purified using oligo (dT) magnetic beads to isolate poly(A) mRNA. Fragmentation buffer was added for fragmenting mRNA to short fragments, and the short fragments were taken as templates to synthesize the first-strand cDNA and reverse transcriptase. DNA polymerase I and RNase H were used to synthesize the second-strand cDNA. The short fragments were conducted with end repair and adapter ligation. These products were purified and enriched with PCR, followed by 15 cycles to create the final cDNA library. The library was sequenced to obtain short sequences of 90 bp from paired-end on an Illumina HiSeq 4000 sequencing platform.
P. octandrus total RNA was extracted from submerged and floating leaves. 10 µg of total RNA was purified using oligo (dT) magnetic beads to isolate poly(A) mRNA. Fragmentation buffer was added for fragmenting mRNA to short fragments, and the short fragments were taken as templates to synthesize the first-strand cDNA and reverse transcriptase. DNA polymerase I and RNase H were used to synthesize the second-strand cDNA. The short fragments were conducted with end repair and adapter ligation. These products were purified and enriched with PCR, followed by 15 cycles to create the final cDNA library. The library was sequenced to obtain short sequences of 90 bp from paired-end on an Illumina HiSeq 4000 sequencing platform.