The expression of AHR and HDAC8 mRNA in hepatoma cells and cells from cancer patients were quantified using the SYBR Green Quantitative RT-PCR kit (Invitrogen) as described previously. Total RNA was extracted from the tumor mass using TRIzol reagent (Invitrogen) and transcribed into cDNA (Invitrogen) for PCR amplification using a 7900HT Thermocycler (Applied Biosystems Inc.). All procedures and data analyses were performed according to the manufacturer's instructions. Data on cells transfected with an empty pEGFP vector and samples from healthy subjects and drug-treated patients were analyzed and compared. All data are expressed as the mean ± SD of at least three experiments.
Sybr green quantitative rt pcr kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The SYBR Green Quantitative RT-PCR kit is a reagent system designed for the detection and quantification of RNA targets using real-time reverse transcription polymerase chain reaction (RT-PCR) technology. The kit includes all the necessary components, including a SYBR Green-based detection dye, to perform quantitative RT-PCR analysis.
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5 protocols using sybr green quantitative rt pcr kit
1
Quantifying AHR and HDAC8 Expression
2
Quantifying IDO1, TDO2, and AHR mRNA in Leukemia Cells
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The expressions of IDO1, TDO2, and AHR mRNA in leukemia cells from cancer patients were quantified using the SYBR Green Quantitative RT-PCR kit (Invitrogen) as described previously. The total RNA was extracted from the tumor mass using TRIzol reagent (Invitrogen) and then transcribed into cDNA (Invitrogen) for PCR amplification on a 7900HT Thermocycler (Applied Biosystems Inc.). All procedures and data analysis were performed according to the manufacturer’s instructions. All data are expressed as the mean ± SD of at least 3 experiments.
3
Analysis of Gene Expression Profiles
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The ISX, cyclin D1, RB, p53 and E2F1 mRNA expression from hepatoma cells and tumor patients was detected by SYBR Green Quantitative RT-PCR kit (Invitrogen) as previous described [41 (link)]. The total RNA was extracted from the tumor mass with Trizol reagent (Invitrogen), and then transcribed into cDNA (Invitrogen) for PCR amplification using an ABI 7900HT Thermocycler. All reactions and data analyses were performed according to the manufacturer's instructions. pEGFP/c1 vector-transfected cells and samples from normal subjects and drug treated patients were analyzed for comparison. All of experiments represents as means±SD and repeated at least three times.
4
Quantifying SPZ1 and TWIST1 mRNA in Hepatoma
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The expressions of SPZ1 and TWIST1 mRNA in hepatoma cells and cells from cancer patients were quantified using the SYBR Green Quantitative RT-PCR kit (Invitrogen, Carlsbad, CA, USA) as described previously. The total RNA was extracted from the tumor mass using TRIzol reagent (Invitrogen) and then transcribed into cDNA (Invitrogen) for PCR amplification on a 7900HT Thermocycler (Applied Biosystems-Thermo Fisher Scientific, Walham, MA, USA). All procedures and the data analysis were performed according to the manufacturers’ instructions. All the data are expressed as the mean±s.d. of at least three experiments.
5
Quantifying ISX, BRD4, and PCAF mRNA in Lung Cancer
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The expression of ISX, BRD4, and PCAF mRNA in lung cancer cells and cells from cancer patients was quantified using an SYBR Green Quantitative RT–PCR kit (Invitrogen) as described previously. Total RNA was extracted from tumor mass using TRIzol reagent (Invitrogen) and then transcribed into cDNA (Invitrogen) for PCR amplification using a 7900HT Thermocycler (Thermo Fisher Scientific, Waltham, MA, USA). All procedures and data analysis were performed according to the manufacturers’ instructions. The cells were transfected with an empty pEGFP vector, and samples from healthy subjects and drug‐treated patients were analyzed for comparison. All data are expressed as mean ± SD of at least three experiments.
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