Total cellular RNAs were isolated by using TRIzol reagent (Life Technologies). cDNAs were prepared using Honor II 1st Strand cDNA Synthesis Supermix for qPCR (Novogene, Tianjin, China). mRNA expression levels were detected by Unique Aptamer Green Master Mix (Novogene, Tianjin, China) on LightCycler 96 Detection System (Roche, Basel, Switzerland). Detection of GAPDH mRNA expression was used for normalization. Primers used in this study are provided in Supplementary Data .
Unique aptamer green master mix
Manufactured by Novogene
Sourced in China
Unique Aptamer Green Master Mix is a ready-to-use solution for real-time PCR applications. It contains all the necessary components, including a proprietary aptamer-based hot-start Taq DNA polymerase, optimized buffer, and fluorescent dye for detecting amplification.
Automatically generated - may contain errors
Lab products found in correlation
Lightcycler 96 detection system, by Roche (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Hoxa9, by Abcam (1 mentions)
Isotype igg, by Merck Group (1 mentions)
Ez chip chromatin immunoprecipitation kit, by Merck Group (1 mentions)
Transzol reagent, by Transgene (1 mentions)
Transscript uni all in one first strand cdna synthesis supermix for qpcr, by Transgene (1 mentions)
3 protocols using unique aptamer green master mix
1
Quantitative Analysis of Gene Expression
2
Chromatin Immunoprecipitation Assay Protocol
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Chromatin immunoprecipitation (ChIP) was done by using EZ ChIP Chromatin Immunoprecipitation kit (Cat. 17-371, Merck Millipore, Burlington, MA, USA). 5 μg antibodies against HOXA9 (Abcam, Cambridge, MA, USA) or isotype IgG (Merck Millipore, Burlington, MA, USA) was used. qRT-PCR was done using Unique Aptamer Green Master Mix (Novogene, Tianjin, China). Relative enrichment was calculated by normalizing to IgG immunoprecipitation. Primers used in this study are listed in Supplementary Data 2 .
3
Extraction and Quantification of mRNA
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Total RNAs from cells were extracted using TransZol reagent (TransGen Biotech Co., Ltd.) according to the manufacturer’s instructions. Reverse transcription was performed using TransScript Uni All-in-One First-Strand cDNA Synthesis SuperMix for qPCR (One-Step gDNA Removal) (TransGen Biotech, AU341). The mRNA expression levels were detected using Unique Aptamer Green Master Mix (Novogene, Tianjin, China) on LightCycler 96 Detection System (Roche, Basel, Switzerland). Detection of GAPDH mRNA expression was used for normalization. Primers used in this study are listed in Supplementary Table S1 .
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