Pgem t easy vector system 1

Manufactured by Thermo Fisher Scientific

The PGEM-T Easy Vector System I is a DNA cloning vector system used for the rapid cloning of PCR products. It provides a convenient and efficient way to insert PCR amplified DNA fragments into a plasmid vector for further analysis and manipulation.

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Lab products found in correlation

Axiophot light microscope, by Zeiss (2 mentions) Dig rna labeling kit, by Roche (1 mentions) Dig rna labelling kit, by Roche (1 mentions)

2 protocols using pgem t easy vector system 1

In Situ Hybridization of Brizantha Ovaries

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Semi-thin Sects. (3.5 μm) of ovaries at megasporogenesis of sexual and apomictic B. brizantha were hybridized with an 866 bp PCR fragment of BbrizIPT9, previously cloned into pGEM-T Easy Vector System I (Invitrogen) and used as template for sense and antisense probes, with SP6 and T7 polymerases respectively. A DIG RNA labeling kit (Roche) was used according to the manufacturer’s protocol. The sample preparation and in situ hybridization procedures were carried out as previously described [13 (link), 17 (link), 18 (link), 42 ]. The hybridized section slides were observed using a Zeiss Axiophot light microscope.

Ferreira L.G., Dusi D.M., Irsigler A.S., Gomes A.C., Florentino L.H., Mendes M.A., Colombo L, & Carneiro V.T. (2023). Identification of IPT9 in Brachiaria brizantha (syn. Urochloa brizantha) and expression analyses during ovule development in sexual and apomictic plants. Molecular Biology Reports, 50(6), 4887-4897.

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RNA in situ Hybridization of BbrizGID1 in Ovary

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The RNA probe was synthesized using the DIG RNA labelling kit (Roche) according to the manufacturer's protocol. The same PCR fragment of 300 bp from BbrizGID1 located in the 3′ region, flanked by positions 1334-1633 of the nucleotide sequence, was used as a probe. The BbrizGID1 fragment was cloned into pGEM-T Easy Vector System I (Invitrogen) and used as a template for in vitro transcription with SP6 and T7 polymerases, used as sense and antisense probes, respectively. In situ hybridization was performed in semi-thin sections of 3.5 μm of ovaries at megasporogenesis of sexual and apomictic plants. The sample preparation and in situ hybridization were carried out as previously described (Alves et al. 2007; Silveira et al. 2012) . Hybridized sections were observed with a Zeiss Axiophot light microscope.

Ferreira L.G., de Alencar Dusi D.M., Irsigler A.S.T., Gomes A.C.M.M., Mendes M.A., Colombo L, & de Campos Carneiro V.T. (2018). GID1 expression is associated with ovule development of sexual and apomictic plants. Plant cell reports, 37(2).

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