Plant total rna isolation kit

The total RNA was extracted from anthurium tissues using an RNAprep Pure Plant Kit (Polysaccharides & Polyphenolics-rich) (TIANGEN, Beijing, China). The total RNA from the Arabidopsis leaves and tobacco corolla limbs was extracted using a Plant Total RNA Isolation Kit (FOREGENE, Chengdu, China). The cDNA was synthesized according to the manufacturer’s instructions for the RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific, Waltham, MA, USA).

The Plant Total RNA Isolation Kit (FOREGENE, Chengdu, China) was adopted to collect elephant grass leaves and extract total RNA, followed by reverse transcription with the HiScript III 1st Strand cDNA Synthesis Kit. Gene expression was normalized with elongation factor-1-alpha (EF1α) as the reference gene. qRT-PCR was made with the CFX96 qRT-PCR System and ChamQ Universal SYBR qPCR Master Mix, with the reaction program following the standard protocol of the kit. Through comparing the relative expression level of target genes, the calculation of control treatments was made with the 2 − ΔΔCt approach. Primer Premier 5.0 software was adopted to design quantitative real-time polymerase chain reaction (qRT-PCR) primers, and Table S4 lists the qRT-PCR primer sequences.

A Plant Total RNA Isolation Kit (FOREGENE, China) was used for total RNA extraction from differently treated samples (i.e., drought, salinity, or heat). A RevertAid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, (Waltham, Massachusetts, USA) was used to synthesize purified cDNA with 1 μg of total RNA and oligo primers [53 (link)]. Light Cycler^® 480 II (Mannheim, Roche, Germany) and SYBR Green I Master Mix (Roche, Germany) were used to perform qPCR. The gene-specific primers were designed using the NCBI online software primer designing tool (https://www.ncbi.nlm.nih.gov/tools/primer-blast/) (accessed on 15 January 2023) (Table S2). Each qPCR reaction was conducted as follows: 0.2 μL of cDNA, 5 μL of 0.5 μM gene-specific primer pre-mixture, 10 μL of 2 × SYBR Green Master Mix, and 4.8 μL of water. Actin7 was used as the internal standard to normalize the expression levels for target genes. A melting curve was used to evaluate the specificity of amplification. All experiments had three biological replicates and technical replicates. The 2−ΔCT method was used for data calculation. Graph Pad Prism 9 was used to analyze and graph the expression data.

Wide-type Anabaena PCC 7120, TRS-polA, and CT-dnaENI were cultivated to the log phase in BG11 with 0.3 μmol CuSO₄ and 1 mM theophylline and then washed with BG11 for three times to remove the inducers. Cells were resuspended in 150-ml fresh BG11 to OD₇₅₀ of 0.3 and cultured at the same conditions for 4 days. To prepare the samples, cultures were diluted to the initial concentration in fresh BG11 and 50 ml of samples were harvested at day 0, day 1, day 2, day 3, and day 4, respectively, with two parallel experiments. Total RNA was extracted with Plant Total RNA Isolation Kit (FOREGENE) and reverse transcribed with HiScript Q RT super mix (Vazyme). qRT-PCR was performed using ChamQ SYBR Qpcr Master Mix (Vazyme, Nanjing, China) with specific primers listed in Supplementary Table S2.

Total RNA was isolated using Plant Total RNA Isolation Kit (Foregene, China). PrimeScript™ RT reagent Kit (Takara, Dalian) was used to generate cDNA according to the manufacturer’s instruction. Quantitative real-time PCR (qRT-PCR) was carried out using SYBR Premix Ex Taq™ II (Takara, Dalian) on a CFX96 real-time PCR system (Bio-Rad, United States). The qRT-PCR cycling began with one cycle at 95°C for 2 min, followed by 40 cycles at 95°C for 30 s, 55°C for 30 s, and 72°C for 30 s. The house-keeping gene, NtGAPDH (XM_016643257), was used as the reference standard. Three biological and three technical replicates were performed. Primers used for qRT-PCR analysis are listed in Supplementary Table S1.

The total RNA of 0.8 g of sweet potato tuberous roots and 0.5 g of strawberry samples was extracted using a Plant Total RNA Isolation Kit (Foregene). First-strand cDNA was synthesized from total RNA using Prime Script™ RT Master Mix (Takara). qRT-PCR was conducted using SYBR® Premix Ex Taq™ II (Takara) in a total reaction volume of 20 μl consisting of 150 ng of template cDNA, each primer at 0.2 μM and 10 μl of SYBR® Premix Ex Taq™ II, and the amplification program was as follows: 1 cycle of 95 °C for 10 s followed by 40 cycles of 95 °C for 5 s and 60 °C for 34 s. A strawberry gene, FvTubulin (gene11892), and a sweet potato gene, IbTubulin (Itf04g29110), were used as internal controls. The primers used for RT-qPCR are listed in additional file 1: Table S1.

Plant tissues were sampled and ground immediately in liquid N₂, total RNA was extracted using the Plant Total RNA Isolation Kit (Foregene, China) and reverse-transcribed into cDNA with PrimeScript™ RT reagent Kit (Takara, Dalian). qRT-PCR program was performed using SYBR Premix Ex Taq™ II (Tli RNaseH Plus) (Takara, Dalian) on a CFX96 real-time PCR system (Bio-Rad, USA). The qRT-PCR cycling began with one cycle at 95 °C for 2 min, followed by 40 cycles at 95 °C for 30 s, 55 °C for 30 s, 72 °C for 30 s. The house-keeping gene, NtGAPDH (XM_016643257), was used as the internal control. Three biological and three technical replicates were performed. All primer used for qRT-PCR were listed in Additional file 2: Table S2.