Anterior and posterior thirds of untreated or retinoic acid–treated wing bud tissue were dissected from eight embryos. Total RNA was extracted using TRIzol Reagent (Life Technologies), purified using a PureLink RNA mini kit (Ambion), and cDNA‐prepared using SuperScript II Reverse Transcriptase (Invitrogen). qPCR was performed on an Applied Biosystems StepOne RT‐PCR machine using TaqMan Fast Advanced Master Mix (Thermo Fisher Scientific) and a TaqMan probe and primer set designed against chicken Shh (Gg03338766_m1, Thermo Fisher Scientific); 5 ng cDNA was used per reaction (20‐μl volume) with cycle conditions of 95 °C for 20 sec, followed by 32 cycles of 95 °C for 1 sec and 60 °C for 20 sec. All reactions were carried out in triplicate and normalized against Eukaryotic 18S rRNA Endogenous Control expression (Thermo Fisher Scientific). Standard error mean bars were generated from the triplicate CT values. Unpaired t‐tests measured significance of expression change between appropriate samples. Applied Biosystems StepOne Software V2.3 was used to analyze the data and generate gene expression comparisons.
Eukaryotic 18s rrna endogenous control expression
Manufactured by Thermo Fisher Scientific
The Eukaryotic 18S rRNA Endogenous Control expression is a laboratory tool used to measure the expression levels of the 18S ribosomal RNA (rRNA) gene, which is commonly used as a reference gene in eukaryotic gene expression studies. It provides a consistent and reliable means of normalizing gene expression data across different samples.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions)
Stepone software v2, by Thermo Fisher Scientific (1 mentions)
Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Purelink rna mini kit, by Thermo Fisher Scientific (1 mentions)
Stepone rt pcr machine, by Thermo Fisher Scientific (1 mentions)
Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Direct zol rna kit, by Zymo Research (1 mentions)
2 protocols using eukaryotic 18s rrna endogenous control expression
1
Quantifying Chicken Sonic Hedgehog Expression
2
Quantification of Cyp26b1 Expression
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Ten whole limb buds at 0, 6, 12 and 24 hours were dissected from either quail or chick embryos.
Total RNA was extracted using TRIzol Reagent (Life Technologies), purified using a Directzol RNA kit (Zymo Research) and cDNA prepared using SuperScript III Reverse Transcriptase (Invitrogen). qPCR was performed on an Applied Biosystems StepOne RTPCR machine using SYBR Green Master Mix (Thermo Fisher Scientific) and a primer set for Cyp26b1 was designed against a sequence which was present in both chicken and quails, spanning exon junctions (Thermo Fisher Scientific). 5 ng cDNA was used per reaction (20μl volume) with cycle conditions of 95 °C for 20 sec, followed by 32 cycles of 95 °C for 1 sec and 60 °C for 20 sec. All reactions were carried out in triplicate and average CT values normalized against Eukaryotic 18S rRNA Endogenous Control expression (Thermo Fisher Scientific). Unpaired student t-tests compared the mean relative expression, and measured significance of expression change between appropriate samples. Applied Biosystems StepOne Software V2.3 was used to analyse the data and GraphPad Prism8 used to construct graphs.
Total RNA was extracted using TRIzol Reagent (Life Technologies), purified using a Directzol RNA kit (Zymo Research) and cDNA prepared using SuperScript III Reverse Transcriptase (Invitrogen). qPCR was performed on an Applied Biosystems StepOne RTPCR machine using SYBR Green Master Mix (Thermo Fisher Scientific) and a primer set for Cyp26b1 was designed against a sequence which was present in both chicken and quails, spanning exon junctions (Thermo Fisher Scientific). 5 ng cDNA was used per reaction (20μl volume) with cycle conditions of 95 °C for 20 sec, followed by 32 cycles of 95 °C for 1 sec and 60 °C for 20 sec. All reactions were carried out in triplicate and average CT values normalized against Eukaryotic 18S rRNA Endogenous Control expression (Thermo Fisher Scientific). Unpaired student t-tests compared the mean relative expression, and measured significance of expression change between appropriate samples. Applied Biosystems StepOne Software V2.3 was used to analyse the data and GraphPad Prism8 used to construct graphs.
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