Ultrasec 2100 pro uv visible spectrophotometer

Total RNA of six leaves from three samples in each genotype was isolated as three biological replications using an RNAprep pure Plant RNA Purification Kit (Tiangen Biotech, Beijing, China). The quality and quantity of total RNA were analyzed using an UltrasecTM 2100 pro UV/Visible Spectrophotometer (Amersham Biosciences, Uppsala, Sweden) and by gel electrophoresis. For each genotype, high-quality RNA from three replications was used for cDNA library construction and Illumina deep sequencing.
A total amount of 3 µg RNA per sample was used as input material for the RNA sample preparations. Fifteen cDNA libraries were generated using NEBNext^® Ultra™ RNA Library Prep Kit for Illumina^® (NEB, USA). RNA detection, cDNA library construction and Illumina deep sequencing were performed following the methods by the Beijing Novogene Biological Information Technology Co., Ltd., Beijing, China (http://www.novogene.com). The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia) according to the manufacturer’s instructions. After cluster generation, the library preparations were sequenced on an Illumina Hiseq 4000 platform and 150 paired-end reads were generated.

Branching traits between these two varieties show differences after stem has four nodes, so, we take samples when the forth node just emerge. Tissues including leaf, stem, shoot and root, were dissected from Salvia splendens of ten plants, and collected samples were then immediately frozen and stored in liquid nitrogen prior to further analysis. Total RNA was extracted from these materials using Norgen RNA Purification Kit (Norgen Biotek Co., Ontario, Canada). The quality and quantity of purified RNA were examined using an UltrasecTM 2100 pro UV/Visible Spectrophotometer (Amersham Biosciences, Uppsala, Sweden) and gel electrophoresis. Equal amounts of high-quality RNA from each material were pooled for cDNA synthesis.

All materials were obtained from Chengdu Institute of Biology, Chinese Academy of Sciences, and naturally grown in flowerpots (Zhongbei, Chengdu, Sichuan, China). The voucher of Youngia japonica is Peng2013. All vouchers of samples in our experiments are deposited in the Herbarium of the Chengdu Institute of Biology (CDBI). This species was identified according to Flora of China [1] . The whole plants (above ground parts, including leaves, stems and flowers) were immediately frozen and stored in liquid nitrogen until analysis. Total RNA was extracted from these materials using an EASY spin microRNA Rapid extraction kit (Aidlab Biotechnologies Co., Ltd., China). The quality and quantity of total RNA were analyzed using an Ultrasec TM 2100 pro UV/Visible Spectrophotometer (Amersham Biosciences, Uppsala, Sweden), gel electrophoresis, and Agilent G2939A (Agilent RNA 6000 Nano Kit). Equal quantities of high-quality RNA from each material were pooled for cDNA synthesis.