Recombinant proteins expressed in Escherichia coli were obtained as described previously (Yin et al., 2021 (link)). The 1 μM recombinant protein was infiltrated into N. benthamiana leaves, and total RNA was extracted using the Plant Total RNA Kit (ZomanBio, Beijing). Relative expression of PTI marker genes of N. benthamiana, NbCYP71D20, and NbPTI5, was determined by real-time qPCR as described previously (Yin et al., 2021 (link)). NbACT was used as the internal reference. Primers used for qPCR are listed in Supplementary Table 1 .
Plant total rna kit
Manufactured by ZOMANBIO
Sourced in China
The Plant Total RNA Kit is a laboratory product designed to isolate and purify total RNA from a variety of plant samples. It utilizes a simple and efficient method to extract high-quality RNA suitable for downstream applications such as reverse transcription and gene expression analysis.
Automatically generated - may contain errors
Lab products found in correlation
Lightcycler 480 2, by Roche (1 mentions)
Tb green premix ex taq 2, by Takara Bio (1 mentions)
Primescript rt master mix, by Takara Bio (1 mentions)
Abi quantstudio 6 flex system, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Transscript one step gdna removal and cdna synthesis supermix kit, by Transgene (1 mentions)
Transstart top green qpcr supermix dye 2, by Transgene (1 mentions)
Cfx96 real time system, by Bio-Rad (1 mentions)
Evagreen 2 qpcr mastermix, by Applied Biological Materials (1 mentions)
Lightcycler 480 instrument, by Roche (1 mentions)
All in one rt mastermix, by Applied Biological Materials (1 mentions)
Brightgreen 2x qpcr mastermix no dye, by Applied Biological Materials (1 mentions)
7 protocols using plant total rna kit
1
Quantifying Plant Immune Response in N. benthamiana
2
Quantifying Potato Virus Y Expression Using RT-qPCR
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The total RNA was extracted using a Plant Total RNA Kit (ZOMANBIO, ZP405-1, Beijing, China), according to the manufacturers’ instructions. The first-strand cDNA was synthesized using a five × All-In-One RT MasterMix (with AccuRT Genomic DNA Removal Kit) Reverse Transcription Kit (Applied Biological Materials Inc., Richmond, Canada). The gene and PVY expression was quantified by RT-qPCR using a LightCycler 480 II (Roche Diagnostics, IN, USA) and EvaGreen two × qPCR Master Mix (Applied Biological Materials, Vancouver, Canada). The potato Ef1a gene was used as an internal control for the data normalization [29 (link)]. The expression levels of the genes were calculated using the 2−ΔCq method described by Bio-Rad (Hercules, CA, USA). All of the primers used for RT-qPCR analysis are listed in Table S2 (Supplementary Materials) .
3
Gene Expression Profiling of Phytophthora capsici Infection
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P. capsici hyphae inoculating on N. benthamiana leaves were collected at 0, 1.5, 6, 12, 24 and 36 hr postinoculation. Total RNA was extracted using a Plant Total RNA Kit (ZomanBio) and cDNA was synthesized by PrimeScript RT Master Mix (Takara) according to the manufacturer's instructions. Real‐time PCR was performed using TB Green Premix Ex Taq II (Takara) on the ABI QuantStudio 6 Flex system (Thermo Fisher). The gene‐specific primers used for RT‐qPCR are listed in TableS3 .
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P. capsici hyphae inoculating on N. benthamiana leaves were collected at 0, 1.5, 6, 12, 24 and 36 hr postinoculation. Total RNA was extracted using a Plant Total RNA Kit (ZomanBio) and cDNA was synthesized by PrimeScript RT Master Mix (Takara) according to the manufacturer's instructions. Real‐time PCR was performed using TB Green Premix Ex Taq II (Takara) on the ABI QuantStudio 6 Flex system (Thermo Fisher). The gene‐specific primers used for RT‐qPCR are listed in Table
4
Quantitative RNA Expression Analysis
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Total RNA was extracted from rice tissues with Plant Total RNA Kit (ZOMANBIO, Beijing, China). PrimeScriptTM RT reagent Kit (TaKaRa, Shiga, Japan) was used to perform reverse transcription. qRT-PCR was performed using 2× HQ SYBR qPCR Mix (without ROX) (ZOMANBIO). The qRT-PCR primers used are listed in Table S1 .
5
Quantitative Gene Expression Analysis
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Total RNA was extracted from leaves of transgenic and WT plants according to the manufacturer’s protocol of plant total RNA kit (ZOMANBIO, ZP405-1, Beijing, China). RNA integrity was analyzed by spectrophotometry and 1% agarose gel electrophoresis. 1μg RNA was took for cDNA synthesis using TransScript One-Step gDNA Removal and cDNA Synthesis SuperMix kit (TransGen, Beijing, China) following the manufacturer’s protocol. Real-time RT-PCR analyses were performed using TransStart Top Green qPCR SuperMix (+Dye II) (TransGen, Beijing, China). The three-step method was performed as follows: 94 °C 30 s; 40 cycles of 94 °C for 5 s, 58 °C for 15 s and 72 °C for 34 s. The Actin gene was used as an internal control. After the amplification process, the relative quantification of gene expression and statistical analysis were calculated using the 2−ΔΔCT [54 (link)]. All primers used for qRT-PCR are shown in Table S1 .
6
Quantifying Gene Expression in Plants
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Total RNA was extracted using the Plant Total RNA kit (Zomanbio) in accordance with the manufacturer's instructions. First‐strand cDNA was synthesized with 1 μg of RNA following the manufacturer's instructions of the All‐In‐One 5× RT MasterMix kit (Applied Biological Materials). Real‐time qPCR was performed using EvaGreen 2× qPCR MasterMix (Applied Biological Materials) and a CFX96 Real‐Time system (Bio‐rad). PCR conditions were 95°C for 15 min, followed by 40 cycles of 95°C for 15 s, 60°C for 30 s, and 72 °C for 30 s. Gene expression levels were calculated by a 2−ΔΔCt method, as described previously (Livak & Schmittgen, 2001 (link)). At least three independent biological replicates of each treatment were performed, and three technical replicates were performed for each biological replicate. The potato gene Stef1a (XM_006343394) (Nicot et al., 2005 (link)) and tobacco gene Ntactin (XM_016658880) were used as internal reference genes. All primers used in RT‐qPCR are listed in Table S2 .
7
Quantitative RNA Expression Analysis in Potato
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Total RNA was extracted according to the manufacturer’s protocol (Plant Total RNA Kit; ZP405-1; ZOMANBIO, Beijing, China). Monitoring the degradation and contamination of the total RNA samples was by a 1% agarose gel electrophoresis. Reverse transcription into cDNA was performed using 5 × All-In-One RT MasterMix (ABM, Richmond, BC, Canada).
Quantitative reverse transcription PCR was performed with Roche LightCycler 480 Instrument. The program was set according to BrightGreen 2X qPCR MasterMix-No Dye (ABM, Richmond, BC, Canada). The potato ef1α gene (GenBank accession: AB061263) was used as an internal reference gene [32 (link)]. Gene expression levels were calculated via the 2−ΔCq method (http://www.bio-rad.com/zh-cn/applications-technologies/real-time-pcrexperimental-design (accessed on 4 June 2022). The primer sequence of qRT-PCR is presented in Table S2 .
Quantitative reverse transcription PCR was performed with Roche LightCycler 480 Instrument. The program was set according to BrightGreen 2X qPCR MasterMix-No Dye (ABM, Richmond, BC, Canada). The potato ef1α gene (GenBank accession: AB061263) was used as an internal reference gene [32 (link)]. Gene expression levels were calculated via the 2−ΔCq method (
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