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To analyze implantation, Kit W-sh/W-sh females (C57BL/6J background) were mated with BALB/c males, because allogeneic matings represent natural, biologically relevant combinations compared with, for example, syngeneic ones.
Homozygous hST3Gal-I female mice on a pure C57Bl/6 background were mated with FVB male mice heterozygous for the polyomavirus middle T antigen driven by the mouse mammary tumor virus (MMTV-PyMT mice (Guy et al. 1992)) promoter to obtain the ST3Gal-I/PyMT mice.
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Bmi1+/−mice (129Ola/FVB/N hybrid background) that were backcrossed 10 12 times onto a C57BL/6J background were mated to generate Bmi1−/−, Bmi1+/− and WT mice (littermates) genotyped by PCR, as described previously.
To assess the effects of genetic background on penetrance of the PPCD1 phenotype, PPCD1 mice on the DBA/2J background (N8 generation) were mated with either FVB/N or C57BL/6J (B6) mice.
To introgress Q40-YFP transgene into the wild backgrounds, Q40Bristol males were mated with DR1350, JU258(Madeira), or CB4856(Hawaii) hermaphrodites, then five to seven F1 fluorescent hermaphrodites were mated with wild males, and this two-step cycle was repeated until over 35 generations of backcrosses to the wild background were reached.
Plg −/+ mice backcrossed into an FVB/n background for 20 generations (Panum Institute, Copenhagen, Denmark) were mated with MMP9 −/+ mice that were backcrossed into an FVB/n background for 15 generations, to produce the F1 generation of MMP9 −/+:Plg −/+ mice.
F1 heterozygous mice were mated with FlpE transgenic "deleter" mice [Tg ACTFLPe 9205Dym, C57Bl/6 background] to remove the Neo cassette from the targeted locus and obtain the F2 generation.
In brief, DA females were mated with male offspring selected for PVG alleles within Eae23 and against PVG background contamination at 96 microsatellite markers equally spaced throughout the genome (20 centiMorgan intervals).
Resulting chimeras were mated with Swiss Black mice, and the targeted stocks maintained on a mixed 129/Swiss Black background.
Chimeras were mated with C57BL/6 mice, and F1 progeny carrying the transgene were backcrossed 6 10 times onto the C57BL/6 background.
Male chimeras were mated with C57BL6/J females to produce F1 breeders and experiments were performed on N2F1 mice (>83% C57Bl6 background; designated VGFR or Vgf R -/-) from two separate clones with similar results.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com