Total rna extraction reagent

Manufactured by Transgene

Sourced in China

The Total RNA Extraction Reagent is a solution designed for the isolation and purification of total RNA from a variety of biological samples. This reagent utilizes a guanidinium thiocyanate-phenol-chloroform extraction method to effectively isolate RNA while maintaining its integrity.

Automatically generated - may contain errors

Lab products found in correlation

Hiscript 2 first strand cdna synthesis kit, by Vazyme (2 mentions) Lightcycler 480, by Roche (2 mentions)

2 protocols using total rna extraction reagent

Evaluating Nodulation Candidate Genes via qRT-PCR

Cited 1 time

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To verify whether the candidate genes were involved in nodulation, total RNA was extracted with a total RNA extraction reagent (Transgene Biotech Co, Beijing, China). First-strand cDNA was synthesized by reverse transcription with the HiScript^® II First-strand cDNA Synthesis Kit (Vazyme Biotech, Nanjing, China). The qRT-PCR assay was completed with the Roche LightCycler 480 quantitative PCR instrument. The qRT-PCR primers were designed based on the locus information in Phytozome, and GmUKN1 (Glyma.12G020500) was used as a reference gene for normalizing the target gene expression levels (Hu et al., 2009 (link)). The qRT-PCR program was as follows: pre-denaturation at 95°C for 30 s and then 40 cycles of release and an annealing temperature of 58°C for 10 s. The qRT-PCR assay was completed with three biological replicates, each comprising three technical replicates, to increase the accuracy of the data.

Zou J., Zhang Z., Yu S., Kang Q., Shi Y., Wang J., Zhu R., Ma C., Chen L., Wang J., Li J., Li Q., Liu X., Zhu J., Wu X., Hu Z., Qi Z., Liu C., Chen Q, & Xin D. (2020). Responses of Soybean Genes in the Substituted Segments of Segment Substitution Lines Following a Xanthomonas Infection. Frontiers in Plant Science, 11, 972.

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Quantifying Nodulation-related Gene Expression

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To determine whether the candidate genes were involved in nodulation, total RNA was extracted using total RNA extraction reagent (Transgene Biotech Co.). Reverse transcription was performed with the HiScript II First-strand cDNA synthesis kit (Vazyme Biotech). The qRT-PCR analyses were conducted using a Roche LightCycler 480 quantitative PCR instrument. Primers were designed based on locus information in Phytozome, and GmUKN1 (Glyma.12g020500) was used as a reference gene for relative quantification of target gene transcript levels (Hu et al. 2009) . The qRT-PCR program was as follows: predenaturation at 95°C for 30 s, and 40 cycles of release and annealing at 58°C for 10 s. Three biological replicates and three technical replicates were used to improve the accuracy of the results.

Shi Y., Zhang Z., Wen Y., Yu G., Zou J., Huang S., Wang J., Zhu J., Wang J., Chen L., Ma C., Liu X., Zhu R., Li Q., Li J., Guo M., Liu H., Zhu Y., Sun Z., Han L., Jiang H., Wu X., Wang N., Zhang W., Yin Z., Li C., Hu Z., Qi Z., Liu C., Chen Q, & Xin D. (2020). RNA Sequencing-Associated Study Identifies GmDRR1 as Positively Regulating the Establishment of Symbiosis in Soybean. Molecular plant-microbe interactions : MPMI, 33(6).

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