To confirm the expression of FlaB mRNA in vivo, tumor tissues of mice were collected 24 h after l -arabinose injection and treated with RNAwait (Biosharp, Hefei, China). We extracted total RNA from the tumors using TRIzol reagent (Tiangen, Beijing, China) and reverse transcribed it into cDNA using a SuperScript II cDNA Synthesis Kit (Takara Biotechnology, Beijing, China). The qPCR was carried out with SYBR Green (Takara Biotechnology). The 16S gene of P. mirabilis was used as a housekeeping gene.48 (link) The primers used are listed in Table S1 .
Superscript 2 cdna synthesis kit
Manufactured by Takara Bio
Sourced in China, Japan
The SuperScript II cDNA Synthesis Kit is a laboratory reagent used for the reverse transcription of RNA into complementary DNA (cDNA). The kit contains the necessary components, including the SuperScript II Reverse Transcriptase enzyme, to efficiently convert RNA into single-stranded cDNA, which can then be used for various downstream applications.
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Lab products found in correlation
3 protocols using superscript 2 cdna synthesis kit
1
In vivo FlaB mRNA Expression Analysis
2
Quantifying Protein Translation Levels
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To detect protein translation levels, total RNA was isolated by TriZOL reagent (Invitrogen) and then reverse-transcribed using the SuperScript II cDNA Synthesis Kit (TAKARA). RT-qPCR was performed by using QuantStudio TM 6 Flex SYBR® Green Reagents system (Applied Biosystems, United States). Each sample was analyzed in triplicate. The internal control used for calibration of gene expression levels between samples was GAPDH mRNA. Analysis change between resistance strains and susceptible strain were calculated using −ΔΔCT. The primers used for PCR are shown in Table 1 .
3
Quantifying Gene Expression in Tumors
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Total RNA was isolated from tumors using Trizol reagent (Beyotime). RNA was reverse transcribed using the SuperScript II cDNA Synthesis Kit (Takara, Japan), and real-time quantitative PCR (RT-qPCR) was performed using SuperReal PreMix Plus (TIANGEN, China). The primers are listed in the Table 1 .
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