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Open Access Highly Accessed Methodology article

Transposon-mediated BAC transgenesis in zebrafish and mice

Maximiliano L Suster1, Kenta Sumiyama23 and Koichi Kawakami13*

Author Affiliations

1 Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan

2 Division of Population Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan

3 Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), 1111 Yata, Mishima, Shizuoka 411-8540, Japan

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BMC Genomics 2009, 10:477  doi:10.1186/1471-2164-10-477

Published: 16 October 2009

Additional files

Additional file 1:

Tol2-mediated BAC excision in early zebrafish embryos. Transient in vivo assay of Tol2-mediated BAC excision in zebrafish embryos, including DNA microinjection procedure and demonstration of iTol2 cassette excision by PCR.

Format: TIFF Size: 2.5MB Download file

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Additional file 2:

Table S1. Survival and Tol2-mediated BAC integration efficiency upon microinjection in mice. Number of surviving and PCR positive embryos for the transgene were counted after injection of BAC DNA with or without transposase mRNA in mouse oocytes.

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Additional file 3:

Minimal requirements for Tol2-mediated BAC transgenesis in mouse oocytes. Schematic of Tol2-BAC microinjection into the mouse oocyte and PCR genotyping of founder mice injected with or without transposase mRNA into either cytoplasm or pronucleus or both.

Format: TIFF Size: 2.8MB Download file

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Additional file 4:

Estimation of Tol2-BAC copy number in transgenic zebrafish and mice. Semi-quantitative PCR on BAC transgenic zebrafish and mouse genomic DNAs. PCR gel band intensities generated from transgenic DNAs were compared to those of control BAC DNA standards to produce an estimate of copy number.

Format: TIFF Size: 2.6MB Download file

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Additional file 5:

Tol2-mediated excision of a 120 kb BAC clone in zebrafish embryos. Schematic of a 120 kb zebrafish BAC clone containing an iTol2 cassette and detection of its excision in vivo by PCR.

Format: TIFF Size: 3.1MB Download file

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Additional file 6:

Figure legends. Detailed legends of additional files

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