Pwd phj

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The PWD/PhJ is a laboratory equipment product. It serves a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach. Information about the intended use of this product is not available.

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7 protocols using pwd phj

Genotyping and Breeding of Consomic Mice

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C57BL/6J (B6), PWD/PhJ (PWD), and B6.Chr^PWD consomic mice were purchased from Jackson Laboratories (Bar Harbor, ME, USA), then bred and housed in a single room within the vivarium at the Larner College of Medicine at the University of Vermont for five or more generations. The experimental procedures used in this study were approved by the Animal Care and Use Committee of the University of Vermont.
To ensure their correct identity and to enhance rigor and reproducibility of these studies, B6.ChrPWD consomic mice were subjected to genome-wide SNP genotyping using DartMouse genotyping services (Dartmouth College, NH, USA), as previously described by us ^{22 (link)}. All mice used in this study were of the expected genotypes, with the following exception. Chr17S^PWD mice were found to carry a homozygous B6-derived interval between 30 and 45 Mb on Chr17, encompassing H-2. Chr17F^PWD mice with the full PWD-derived Chr17 were generously provided by Dr. Jiri Forejt (Institute of Molecular Genetics of the ASCR, Czech Republic) and housed in the vivarium at UVM for two or more generations prior to experimentation.

Lahue K.G., Lara M.K., Linton A.A., Lavoie B., Fang Q., McGill M.M., Crothers J.W., Teuscher C., Mawe G.M., Tyler A.L., Mahoney J.M, & Krementsov D.N. (2020). Identification of novel loci controlling inflammatory bowel disease susceptibility utilizing the genetic diversity of wild-derived mice. Genes and immunity, 21(5), 311-325.

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Genotyping and Breeding of Consomic Mice

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Mouse Strain Characterization for Immunology

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A total of 23 mouse strains (129X1/SvJ, A/J, AKR/J, B10.S-H2s/SgMcdJ (B10.S), BALB/cJ, BPL/1J, BPN/3J, C3H/HeJ, C57BL/6J, C57BL/10J, CBA/J, CZECHII/EiJ, DBA/1J, DBA/2J, FVB/NJ, JF1/MsJ, MOLF/EiJ, MRL/MpJ, NOD/ShiLtJ, NU/J, PWD/PhJ, PWK/PhJ, SJL/J and SWR/J were purchased from the Jackson Laboratory (Bar Harbor, ME). All mice, including B10.S-Histh^SJL and B10.S-Histh^SJL ISRC lines, were generated and maintained under specific pathogen-free conditions in the vivarium of the Given Medical Building at the University of Vermont according to National Institutes of Health guidelines. All animal studies were approved by the Institutional Animal Care and Use Committee of the University of Vermont.

Tyler A.L., Raza A., Krementsov D.N., Case L.K., Huang R., Ma R.Z., Blankenhorn E.P., Teuscher C, & Mahoney J.M. (2019). Network-Based Functional Prediction Augments Genetic Association To Predict Candidate Genes for Histamine Hypersensitivity in Mice. G3: Genes|Genomes|Genetics, 9(12), 4223-4233.

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Imprinting Analysis of Aebp2 in Mice

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Embryos with the two different stages, 10.5 and 14.5, were obtained through timed mating between male and female mice of the C57BL/6J strain. The harvested embryos were used for DNA isolation and methylation analyses. For imprinting test, male and female mice of the C57BL/6J strain were bred individually with female and male mice of the PWD/PhJ strain (Jackson Lab, Stock No. 004660). One-day-old F1 pups from these crosses were sacrificed and used for isolating DNA and RNA. The isolated DNA was treated with the bisulfite conversion protocol for DNA methylation analyses. To test the mono-allelic expression (imprinting) of Aebp2, the isolated RNA was reverse-transcribed using the SuperScript III First-Strand Synthesis System (Invitrogen). The resulting cDNA was used as a template for PCR designed to amplify the transcribed region of Aebp2 spanning Exon1a through 2. The RT-based imprinting test was conducted in three individuals (littermates) from each reciprocal cross. One of these individuals from the reciprocal cross was selected for bisulfite sequencing-based imprinting test.
All the detailed information regarding the primer sets for RT-PCR are available in S1 Table.

Kim H., Bakshi A, & Kim J. (2015). Retrotransposon-Derived Promoter of Mammalian Aebp2. PLoS ONE, 10(4), e0126966.

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Meiotic Recombination Protocol in Mice

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Experiments conformed to the US Office of Laboratory Animal Welfare regulatory standards and were approved by the Memorial Sloan Kettering Cancer Center Institutional Animal Care and Use Committee. Mice were maintained on regular rodent chow with continuous access to food and water until euthanasia by CO₂ asphyxiation prior to tissue harvest. Previously described Spo11 (Baudat et al. 2000 (link)), Atm (Barlow et al. 1996 (link)), and Exo1^DA (Zhao et al. 2018 (link)) mutations were maintained on a congenic B6 strain background. The Dmc1 mutation (Pittman et al. 1998 (link)) was maintained on a mixed (129/SV and B6) background. B6xPWD F1 hybrid mice (semifertile) were generated by crossing B6 (stock no. 000664) female mice and PWD/PhJ (stock no. 004660) male mice obtained from the Jackson Laboratory.

Yamada S., Hinch A.G., Kamido H., Zhang Y., Edelmann W, & Keeney S. (2020). Molecular structures and mechanisms of DNA break processing in mouse meiosis. Genes & Development, 34(11-12), 806-818.

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Mouse Diversity Panel Protocol

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Thirty-six male inbred mouse strains (129S1/SvImJ, 129X1/SvJ, A/J, AKR/J, BALB/cByJ, BTBR T⁺Itpr3^tf/J, BUB/BnJ, C3H/HeJ, C57BLKS/J, C57BL/6J, C57BR/cdJ, C58/J, CBA/J, CZECHII/EiJ, DBA/2J, FVB/NJ, I/LnJ, KK/HiJ, LG/J, LP/J, MA/MyJ, NOD/LtJ, NON/LtJ, NZB/BINJ, NZO/HiLtJ, NZW/LacJ, PERA/EiJ, PL/J, PWD/PhJ, PWK/PhJ, RIIIS/J, SEA/GnJ, SJL/J, SM/J, SWR/J, and WSB/EiJ), aged 10–12 weeks, were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). This panel of isogenic mice was chosen based on priority strains from the Mouse Diversity Panel.26 (link) Four mice were used per strain. Male mice were housed four per cage in polycarbonate cages on a 12-hour light/dark cycle (lights on at 7 am), with access to food and water ad libitum. All procedures were approved by the Institutional Animal Care and Use Committee and followed the guidelines set forth by the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals.

Frick A., Fedoriw Y., Richards K., Damania B., Parks B., Suzuki O., Benton C.S., Chan E., Thomas R.S, & Wiltshire T. (2015). Immune cell-based screening assay for response to anticancer agents: applications in pharmacogenomics. Pharmacogenomics and Personalized Medicine, 8, 81-98.

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Genetic Diversity of Mouse Strains

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129S1/SvImJ, A/J, AKR/J, BALB/cByJ, BTBRT⁺tf/J, BUB/BnJ, C3H/HeJ, C57BL/10J, C57BL/6J, C57BLKS/J, C57BR/cdJ, C57L/J, CAST/EiJ, CBA/J, DBA/2J, FVB/NJ, KK/HlJ, LP/J, MRL/MpJ, NOD.B10Sn-H2^b/J (a congenic strain with the NOD genetic background but with a histocompatibility locus from a diabetes-resistant strain), NON/ShiLtJ, NZO/HlLtJ, NZW/LacJ, P/J, PL/J, PWD/PhJ, RIIIS/J, SJL/J, SM/J, SWR/J, and WSB/J were obtained from The Jackson Laboratory (Bar Harbor, ME). Three of the strains were not evaluated at all time points because of lymphoma development (AKR/J), lack of sufficient mice for analysis (CAST/EiJ), or self-mutilations resulting in euthanasia for humane purposes (SJL/J). (9 , 10 (link)) Taken together the strains selected were genetically diverse, which was estimated by SNP genotyping of over 100 strains into 7 distinct genetic groups, so that there were 3–7 strains representing each of the groups.(11 (link))

Linn S.C., Mustonen A.M., Silva K.A., Kennedy V.E., Sundberg B.A., Bechtold L.S., Alghamdi S., Hoehndorf R., Schofield P.N, & Sundberg J.P. (2018). Nail Abnormalities Identified in an Aging Study of 30 Inbred Mouse Strains. Experimental dermatology, 28(4), 383-390.

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