Total RNA was extracted and purified using Aurum total RNA mini kit (Bio-Rad). After NanoDrop RNA quantification (Thermo Scientific), reverse transcription was done with the GoScript Reverse cDNA Synthesis kit (Promega). Quantitative PCR (qPCR) reactions were performed in a final volume of 10 μl using VeriQuest SYBR Green qPCR Master Mix (Affymetrix) with the PikoReal Real-Time PCR System (Thermo Scientific): preincubation at 95°C for 7 min, 40 cycles at 95°C for 15 s, 60°C for 60 s followed by melting curves. Primers are listed in Supplementary Table 3 . Levels of CYCLIN-DEPENDENT KINASE A-1 (CDKA-1) and UBIQUITIN 10 (UBQ10) were used as normalization factors. Data present the transcript levels of three pooled organs, each sample being assessed by three technical replicates.
Goscript reverse cdna synthesis kit
Manufactured by Promega
The GoScript Reverse cDNA Synthesis kit is a tool for reverse transcribing RNA into complementary DNA (cDNA). It provides the necessary components, including a reverse transcriptase enzyme, buffer, and oligo(dT) primers, to facilitate the conversion of RNA into first-strand cDNA.
Automatically generated - may contain errors
Lab products found in correlation
Aurum total rna mini kit, by Bio-Rad (2 mentions)
Veriquest sybr green qpcr master mix, by Thermo Fisher Scientific (1 mentions)
Pikoreal real time pcr system, by Thermo Fisher Scientific (1 mentions)
Janus automated workstation, by PerkinElmer (1 mentions)
Rna clean and concentrator 5, by Zymo Research (1 mentions)
Lightcycler 480, by Roche (1 mentions)
Lightcycler 480 sybr green 1 master mix, by Roche (1 mentions)
2 protocols using goscript reverse cdna synthesis kit
1
RNA Extraction and qPCR Analysis
2
Quantitative Real-Time PCR Analysis
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Total RNA was extracted from whole root organs (Aurum Total RNA Mini Kit, Biorad, USA) and submitted to a second purification step (RNA Clean and Concentrator-5, Zymo Research). Reverse transcription was done with the GoScript Reverse cDNA Synthesis kit (Promega). The Janus Automated Workstation (PerkinElmer) handled all pipetting for PCRs in a final volume of 5 µl using LightCycler 480 SYBR Green I Master mix (Roche). Quantitative PCRs (qPCRs) were performed with the LightCycler 480 (Roche): pre-incubation at 95 °C for 10 min, 45 cycles of 95 °C for 30 s, 60 °C for 60 s followed by melting curves. Primers are listed in Supplementary Table S1. Levels of ACTIN 2 (ACT2) and UBIQUITIN 10 (UBQ10) were used as normalization factors.
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