Protocol detail

Characterization of TCP24 Gene Expression

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The full-length coding sequence of TCP24 was polymerase chain reaction (PCR) amplified and cloned into pBluescript SK. Digoxigenin-labeled sense and antisense probes were synthesized with T7 or T3 RNA polymerase (Roche). Inflorescences from wild type and transgenic plants were pretreated and hybridized as described previously (Liu et al., 2011 (link)). Locked nucleic acid (LNA)-modified probe of miR319a was synthesized and labeled with DIG at the 3′ end and used for in situ hybridization.

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Wang H., Mao Y., Yang J, & He Y. (2015). TCP24 modulates secondary cell wall thickening and anther endothecium development. Frontiers in Plant Science, 6, 436.

Publication 2015

T3 RNA polymerase

Coding sequence Digoxigenin In situ hybridization Inflorescences type Nucleic acid probe Polymerase chain reaction T3 rna polymerase Transgenic plants

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Variable analysis

independent variables

Manipulated: Inflorescences from wild type and transgenic plants

dependent variables

Measured: Expression of TCP24 and miR319a

control variables

Kept constant: Pretreatment and hybridization conditions as described previously (Liu et al., 2011)

controls

Positive control: Not specified
Negative control: Not specified

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