Computational protocol: Overexpression of a novel MADS-box gene SlFYFL delays senescence, fruit ripening and abscission in tomato

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Protocol publication

[…] Total RNA of tomato was extracted using TRIzol (Invitrogen, USA) according to the manufacturer's instructions. Then 1 μg total RNA was used to synthesis first strand cDNA through reverse transcription polymerase chain reaction (M-MLV reverse transcriptase, Takara) with Oligo d(T)18 primer. 1–2 μl cDNA was used to clone the full length of SlFYFL gene with primers SlFYFL-F and SlFYFL-R () through high fidelity PCR (Prime STARTM HS DNA polymerase, Takara). The amplified products were tailed by using DNA -Tailing kit (Takara) and linked with pMD18-T vector (Takara). Positive clones were picked out via Escherichia coli JM109 transformation and confirmed by sequencing (BGI, China). Multiple sequence alignments were performed by DNAMAN version 5.2.2. The phylogenic tree was calculated by MEGA (Molecular Evolutionary Genetics Analysis) version 3.1. […]

Pipeline specifications

Software tools DNAMAN, MEGA
Application Phylogenetics
Organisms Solanum lycopersicum