Paraleonurus sibiricus

R1

sing Agilent 2100 Bioanalyzer ( Agilent RNA 6000 Nano Kit) to do the total RNA sample QC: RNA concentration, RIN value,28S/18S and the fragment length distribution. For plant and fungi samples, we use NanoDropTM to identify the purity of the RNA samples.1)mRNA enrichment: Oligo dT Selection or rRNA depletion;2)RNA fragment and reverse transcription: Fragment the RNA and reverse transcription to double-strand cDNA (dscDNA) by N6 random primer;3)End repair, add A tailing and adaptor ligation:The synthesized cDNA was subjected to end-repair and then was 3 adenylated. Adaptors were ligated to the ends of these 3 adenylated cDNA fragments;4)PCR amplification:The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer;5)Denature and cyclization:Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase;6)Sequencing on BGISEQ-500 platform

{'tissue': 'roots', 'isolate': 'Shantou', 'cultivar': 'Leonurus sibiricus', 'dev_stage': 'maturation period 4', 'geo_loc_name': 'China:Shantou', 'biosamplemodel': 'Plant'}

R2

sing Agilent 2100 Bioanalyzer ( Agilent RNA 6000 Nano Kit) to do the total RNA sample QC: RNA concentration, RIN value,28S/18S and the fragment length distribution. For plant and fungi samples, we use NanoDropTM to identify the purity of the RNA samples.1)mRNA enrichment: Oligo dT Selection or rRNA depletion;2)RNA fragment and reverse transcription: Fragment the RNA and reverse transcription to double-strand cDNA (dscDNA) by N6 random primer;3)End repair, add A tailing and adaptor ligation:The synthesized cDNA was subjected to end-repair and then was 3 adenylated. Adaptors were ligated to the ends of these 3 adenylated cDNA fragments;4)PCR amplification:The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer;5)Denature and cyclization:Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase;6)Sequencing on BGISEQ-500 platform

{'tissue': 'roots', 'isolate': 'Shantou', 'cultivar': 'Leonurus sibiricus', 'dev_stage': 'maturation period 5', 'geo_loc_name': 'China:Shantou', 'biosamplemodel': 'Plant'}

R3

sing Agilent 2100 Bioanalyzer ( Agilent RNA 6000 Nano Kit) to do the total RNA sample QC: RNA concentration, RIN value,28S/18S and the fragment length distribution. For plant and fungi samples, we use NanoDropTM to identify the purity of the RNA samples.1)mRNA enrichment: Oligo dT Selection or rRNA depletion;2)RNA fragment and reverse transcription: Fragment the RNA and reverse transcription to double-strand cDNA (dscDNA) by N6 random primer;3)End repair, add A tailing and adaptor ligation:The synthesized cDNA was subjected to end-repair and then was 3 adenylated. Adaptors were ligated to the ends of these 3 adenylated cDNA fragments;4)PCR amplification:The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer;5)Denature and cyclization:Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase;6)Sequencing on BGISEQ-500 platform

{'tissue': 'roots', 'isolate': 'Shantou', 'cultivar': 'Leonurus sibiricus', 'dev_stage': 'maturation period 6', 'geo_loc_name': 'China:Shantou', 'biosamplemodel': 'Plant'}