We adjusted the concentration of the cDNA template to 100 ng/µL using a trace nucleic acid analysis and prepared the reaction system according to the Power SYBR Green MasterMix Kit (Takara, Kyoto, Japan). The A. westerdijkiae GADPH gene was used as an internal standard [5 (link)]. Gene expression was analyzed using an ABI 7500 real-time quantitative PCR system (Thermo Fisher, Waltham, MA, USA). The amplification primers used for RT-PCR are shown in Table 2 ; the relative expression tests for each gene were repeated three times. Data were processed according to the ABI 7500 v2.0.6 software (Thermo Fisher, Waltham, MA, USA), and relative expression of genes was calculated by the 2−ΔΔCt method.
Power sybr green mastermix kit
Manufactured by Takara Bio
Sourced in Japan
The Power SYBR Green MasterMix Kit is a ready-to-use solution for real-time PCR amplification and detection. It contains SYBR Green I dye, which binds to double-stranded DNA, enabling the quantification of target sequences. The kit provides the necessary components for efficient and reproducible real-time PCR reactions.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using power sybr green mastermix kit
1
Real-Time qPCR Gene Expression Analysis
2
Quantifying PD-L1 Expression via qRT-PCR
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Total RNA of cell lines was extracted using TRIzol and used to synthesize
complementary DNA (cDNA) by reverse transcription through M-MLV (an
RNA-dependent DNA polymerase that can be used in cDNA synthesis with long
messenger RNA templates) and cDNA amplification using the Power SYBR Green
Master Mix kit (Takara, Otsu, Japan). In addition, total RNA for miRNAs was
collected using a High Pure miRNA isolation kit (Roche) and real-time polymerase
chain reaction (RT-PCR) using a TaqMan MicroRNA Reverse Transcription kit (Life
Technologies, Carlsbad, CA, USA). A miScript Primer Assay (QIAGEN) was used for
the miR-191-5p and U6. Data were analyzed using the comparative Ct method (2–ΔΔCt). The procedure of
quantification of PD-L1 expression in RT-PCR was conducted as previously described.21 (link) The primers are listed inSupplementary Table S2 .
complementary DNA (cDNA) by reverse transcription through M-MLV (an
RNA-dependent DNA polymerase that can be used in cDNA synthesis with long
messenger RNA templates) and cDNA amplification using the Power SYBR Green
Master Mix kit (Takara, Otsu, Japan). In addition, total RNA for miRNAs was
collected using a High Pure miRNA isolation kit (Roche) and real-time polymerase
chain reaction (RT-PCR) using a TaqMan MicroRNA Reverse Transcription kit (Life
Technologies, Carlsbad, CA, USA). A miScript Primer Assay (QIAGEN) was used for
the miR-191-5p and U6. Data were analyzed using the comparative Ct method (2–ΔΔCt). The procedure of
quantification of PD-L1 expression in RT-PCR was conducted as previously described.21 (link) The primers are listed in
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