Illumina NovaSeq 6000 paired end sequencing; RNA-sequence to compare the transcriptional profiles of nuclear and plastidic genes in the atnusg and the wild-type
RNA-sequence to compare the transcriptional profiles of nuclear and plastidic genes in the atnusg and the wild-type
Total RNA was isolated from two-week-old wild-type and atnusg mutant young leaves.  For total RNA isolation, frozen sample was ground in liquid nitrogen, and total RNA were extracted using Trizol reagent plus RNeasy mini kit,according to the manufacturerâ€™s instructions  mRNA was enriched using oligo-(dT) primer for nuclear-encoded genes and reverse-transcribed using random hexamers primers. The double-stranded cDNA was purified using AMPure XP beads and treated with End Prep Enzyme Mix to repair both ends and to add a dA-tail in one reaction, followed by T-A ligation to add adaptors to both ends. Size selection of adaptor-ligated DNA was performed using AMPure XP beads, and ~360-bp fragments (with an approximate insert size of 300 bp) were recovered. The library was then constructed and sequenced using an Illumina platform