Rose Petal RNA Extraction and qPCR Analysis

Total RNA was extracted from rose petals by a pBIOZOL kit (BioFlux, BSC55Ml)^{88 (link)}. Genomic DNA contaminants were removed by digestion with RNase-free DNase I (Promega). Reverse transcription was performed with 1 μg total RNA using HiScript II Q Select RT SuperMix kit (Vazyme, Nanjing, China). qPCR was performed on a StepOne Real-Time PCR System (Applied Biosystems, Carlsbad, USA) and a KAPA SYBR FAST qPCR kit (Kapa Biosystems, Wilmington, MA, USA). RhUBI2 was used as an internal control^{86 (link)}.

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Jing W., Gong F., Liu G., Deng Y., Liu J., Yang W., Sun X., Li Y., Gao J., Zhou X, & Ma N. (2023). Petal size is controlled by the MYB73/TPL/HDA19-miR159-CKX6 module regulating cytokinin catabolism in Rosa hybrida. Nature Communications, 14, 7106.

Publication 2023

pBIOZOL kit RNase-free DNase I HiScript II Q Select RT SuperMix kit StepOne Real-Time PCR System KAPA SYBR FAST qPCR kit

Digestion Dnase Genomic Promega Reverse transcription Rnase i

Corresponding Organization :

Other organizations : China Agricultural University, Shenzhen Polytechnic

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expression levels measured by qPCR

control variables

Total RNA extraction method (pBIOZOL kit)
Removal of genomic DNA contaminants (RNase-free DNase I digestion)
Reverse transcription method (HiScript II Q Select RT SuperMix kit)
QPCR method (StepOne Real-Time PCR System, KAPA SYBR FAST qPCR kit)
Internal control gene (*RhUBI2*)

positive controls

None mentioned

negative controls

None mentioned

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