Total RNA was extracted from VP-incubated cells utilizing the TRIzol regent (Invitrogen; Thermo Fisher Scientific, Inc.) using established protocol. cDNA was synthesized using the PrimeScript reverse transcription reagent kit (TIANGEN BIOTECH Co., Beijing, China) by following established guidelines. cDNA was assessed using qPCR on a SuperReal PreMix Plus (SYBR Green) system (TIANGEN BIOTECH Co.). The amplification conditions included: 95 °C for 15 min, 40 cycles of 95 °C for 20 s, 56 °C for 30 s and 68 °C for 30 s. The primer sequences used for qPCR were: YAP forward (F) 5′-TGACCCTCGTTTTGCCATGA-3′ and reverse (R), 5′-GTTGCTG CTGGTTGGAGTTG-3′; CTGF F, 5′-TGGAAGAGAACATTAAGAA GGGCA-3′ and R, 5′-TGCAGCCAGAAAGCTCAAAC-3′; AXL F, 5′-ACCCCAG AGGTGCTAATGGA-3′ and R, 5′-GTGGACTGGCTG TGCTTCC-3′; CYR61 F, 5′-GCAAGGAGCTGGGATTCGAT-3′ and R,5′-ATTCCAAAAACAGGGAGCCG-3′; GAPDH F, 5′-GCACCGTCAAGGCTGAGAAC-3′ and R, 5′-TGGTGAAGACGC CAGTGGA-3′. GAPDH functioned as the internal control. Gene expression was measured utilizing the 2-ΔΔCt method.
Primescript reverse transcription reagent kit
Manufactured by Tiangen Biotech
Sourced in China
The PrimeScript reverse transcription reagent kit is a laboratory product that enables the conversion of RNA into complementary DNA (cDNA) through the process of reverse transcription. The kit includes the necessary reagents and enzymes required for this process.
Automatically generated - may contain errors
5 protocols using primescript reverse transcription reagent kit
1
Quantifying Gene Expression in VP-treated Cells
2
Quantitative Real-Time PCR Expression Analysis
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Total RNA was extracted using TRIzol reagent and treated with DNase. Then, reverse transcription was performed using the PrimeScript reverse transcription reagent kit (TIANGEN, Beijing, China). Quantitative real-time polymerase chain reaction was performed as described previously (He et al., 2020 (link)), using a Light Cycler 480 system (Roche, Shanghai, China), specific primers (Table 2 ), and SYBR Green mix (TIANGEN). The comparative Ct value method was used to quantify mRNA expression relative to GAPDH expression.
3
Quantitative Analysis of Gene Expression
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Total RNA was obtained from VP-incubated cells utilizing the Trizol regent (Invitrogen; Thermo Fisher Scienti c, Inc.) as per established protocol. cDNA was made utilizing the PrimeScript reverse transcription reagent kit (TIANGEN BIOTECH Co., Beijing, China) as per established guidelines. cDNA was assessed through qPCR with a SuperReal PreMix Plus (SYBR Green) system (TIANGEN BIOTECH Co.). The ampli cation conditions include: 95˚C for 15 min, and then 40 cycles of 95˚C for 20 sec, 56˚C for 30 sec and 68˚C for 30 sec. The primer sequences used in qPCR were: YAP forward (F) 5'-TGACCCTCGTTTTGCCATGA-3' and reverse (R), 5'-GTT GCTGCTGGTTGGAGTTG-3'; CTGF F, 5'-TGGAAGAGAACATTAAGAAG GGCA-3' and R, 5'-TGCAGCCAGAAAGCTCAAAC-3'; AXL F, 5'-ACCCC AGAGGTGCTAATGGA-3' and R, 5'-GTGGACTGGCTG TGCTTCC-3'; CYR61 F, 5'-GCAAGGAGCTGGGATTCGAT-3' and R, 5'-ATTCCAAAAACAGGGAGCCG-3'; GAPDH F, 5'-GCACCGTCAAGG CTGAGAAC-3' and R, 5'-TGGTGAAGACGC CAGTGGA-3'. GAPDH functioned as internal control. Gene expression was measured utilizing the 2-ΔΔ Ct method.
4
Quantitative PCR Analysis of Cellular Genes
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Total RNA was obtained from VP-incubated cells utilizing the Trizol regent (Invitrogen; Thermo Fisher Scienti c, Inc.) as per established protocol. cDNA was made utilizing the PrimeScript reverse transcription reagent kit (TIANGEN BIOTECH Co., Beijing, China) as per established guidelines. cDNA was assessed through qPCR with a SuperReal PreMix Plus (SYBR Green) system (TIANGEN BIOTECH Co.). The ampli cation conditions include: 95˚C for 15 min, and then 40 cycles of 95˚C for 20 sec, 56˚C for 30 sec and 68˚C for 30 sec. The primer sequences used in qPCR were: YAP forward (F) 5'-TGACCCTCGTTTTGCCATGA-3' and reverse (R), 5'-GTT GCTGCTGGTTGGAGTTG-3'; CTGF F, 5'-TGGAAGAGAACATTAAGAAG GGCA-3' and R, 5'-TGCAGCCAGAAAGCTCAAAC-3'; AXL F, 5'-ACCCC AGAGGTGCTAATGGA-3' and R, 5'-GTGGACTGGCTG TGCTTCC-3'; CYR61 F, 5'-GCAAGGAGCTGGGATTCGAT-3' and R,5'-ATTCCAAAAACAGGGAGCCG-3'; GAPDH F, 5'-GCACCGTCAAGG CTGAGAAC-3' and R, 5'-TGGTGAAGACGC CAGTGGA-3'. GAPDH functioned as internal control. Gene expression was measured utilizing the 2-ΔΔ Ct method.
5
Quantitative Analysis of Gene Expression
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Total RNA was extracted from VP-incubated cells utilizing the TRIzol regent (Invitrogen; Thermo Fisher Scienti c, Inc.) using established protocol. cDNA was synthesized using the PrimeScript reverse transcription reagent kit (TIANGEN BIOTECH Co., Beijing, China) by following established guidelines. cDNA was assessed using qPCR on a SuperReal PreMix Plus (SYBR Green) system (TIANGEN BIOTECH Co.). The ampli cation conditions included: 95˚C for 15 min, 40 cycles of 95˚C for 20 sec, 56˚C for 30 sec and 68˚C for 30 sec. The primer sequences used for qPCR were: YAP forward (F) 5'-TGACCCTCGTTTTGCCATGA-3' and reverse (R), 5'-GTTGCTGCTGGTTGGAGTTG-3'; CTGF F, 5'-TGGAAGAGAACATTAAGAA GGGCA-3' and R, 5'-TGCAGCCAGAAAGCTCAAAC-3'; AXL F, 5'-ACCCCAG AGGTGCTAATGGA-3' and R, 5'-GTGGACTGGCTG TGCTTCC-3'; CYR61 F, 5'-GCAAGGAGCTGGGATTCGAT-3' and R,5'-ATTCCAAAAACAGGGAGCCG-3'; GAPDH F, 5'-GCACCGTCAAGGCTGAGAAC-3' and R, 5'-TGGTGAAGACGC CAGTGGA-3'. GAPDH functioned as the internal control. Gene expression was measured utilizing the 2-ΔΔCt method.
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