Following total RNA extraction using TRiZOL reagent, cDNA was synthesized using MMuLV Reverse Transcriptase (NEB) with random hexamer primers (Integrated DNA Technologies). Negative (no RT) controls were performed for each target. Quantitative RT-PCR analyses were performed using Brilliant III SYBR Green QPCR master mix (Bioline) with gene-specific primers according to the manufacturer’s protocol and the Applied Bioscience StepOnePlus qRT-PCR machine (Life Technologies). The expression levels were normalized to the endogenous 18S rRNA expression using the delta-delta comparative threshold method (ΔΔCT). Fold changes were determined using the comparative threshold cycle (CT) method (S1 Table ). Efficiencies for primer sets used in this study have been validated in our previous study [1 (link)].
Applied bioscience steponeplus qrt pcr machine
Manufactured by Thermo Fisher Scientific
The Applied Bioscience StepOnePlus is a qRT-PCR (quantitative Reverse Transcription Polymerase Chain Reaction) machine. It is used to amplify and quantify target nucleic acid sequences.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using applied bioscience steponeplus qrt pcr machine
1
Quantitative RT-PCR Analysis of Gene Expression
2
Quantitative RT-PCR Analysis of Fly Transcripts
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Whole flies were homogenized in TRIzol reagent, followed by RNA extraction. cDNA was synthesized using MMulV Reverse Transcriptase (New England Biolab) with random hexamer primers (Integrated DNA Technologies). Negative (no RT) controls were performed for each target. Quantitative RT-PCR analyses were performed using Brilliant III SYBR green QPCR master mix (Agilent) with gene-specific primers according to the manufacturer's protocol and with the Applied Bioscience StepOnePlus qRT-PCR machine (Life Technologies). The expression levels were normalized to the endogenous 18S rRNA expression using the delta-delta comparative threshold method (ΔΔCT). Fold changes were determined using the comparative threshold cycle (CT) method (S1 Table ).
3
Quantitative RT-PCR Analysis of Gene Expression
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Following total RNA extraction using TRiZOL reagent, cDNA was synthesized using MMuLV Reverse Transcriptase (NEB) with random hexamer primers (Integrated DNA Technologies).
Negative (no RT) controls were performed for each target. Quantitative RT-PCR analyses were performed using Brilliant III SYBR Green QPCR master mix (Bioline) with gene-specific primers according to the manufacturer's protocol and the Applied Bioscience StepOnePlus qRT-PCR machine (Life Technologies). The expression levels were normalized to the endogenous 18S rRNA expression using the delta-delta comparative threshold method (ΔΔCT). Fold changes were determined using the comparative threshold cycle (CT) method (Table S1). Efficiencies for primer sets used in this study have been validated in our previous study (1) .
Negative (no RT) controls were performed for each target. Quantitative RT-PCR analyses were performed using Brilliant III SYBR Green QPCR master mix (Bioline) with gene-specific primers according to the manufacturer's protocol and the Applied Bioscience StepOnePlus qRT-PCR machine (Life Technologies). The expression levels were normalized to the endogenous 18S rRNA expression using the delta-delta comparative threshold method (ΔΔCT). Fold changes were determined using the comparative threshold cycle (CT) method (Table S1). Efficiencies for primer sets used in this study have been validated in our previous study (1) .
4
Quantitative RT-PCR for Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following total RNA extraction using TRiZOL reagent, cDNA was synthesized using MMuLV Reverse Transcriptase (NEB) with random hexamer primers (Integrated DNA Technologies). Negative (no RT) controls were performed for each target. Quantitative RT-PCR analyses were performed using Brilliant III SYBR green QPCR master mix (Bioline) with gene-specific primers according to the manufacturer's protocol and with the Applied Bioscience StepOnePlus qRT-PCR machine (Life Technologies). The expression levels were normalized to the endogenous 18S rRNA expression using the delta-delta comparative threshold method (ΔΔCT). Fold changes were determined using the comparative threshold cycle (CT) method (Primer Table ).
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