ENU19 WT:007:b:B6:G3

Abstract

All pedigrees screened in the pipeline will be sourced from mutagenised C57BL/6NCr males. Breeding through to G3 should proceed. The ABS ENU Colony Coordinator will manage the breeding process following the Work Packages supplied annually by APF. G1 mice will be bred to excess. Excess male G1 mice will be reserved for the G1 sequencing project. G2 mice will be restricted to only what is required to establish 2 G3 breeder pairs. The G2 pair is to be disbanded at this stage and the male G1 also reserved for the G1 sequencing project. G3 mice will be bred in two pairs per pedigree. Each pair will produce an average of 16 mice. All weaned G3 mice must be phenotyped by ABS staff according to standard physical and behavioural indicators. This phenotyping will be conducted without the creation of a batch. Each mouse is restrained and its length from nose tip to anus is measured using callipers. The mouse is then weighed and both results recorded on an Excel master sheet that tracks the mouse ID, batch and data. BMI is calculated as weight(kg)/length(m)2 and is also recorded on the spreadsheet. Serum biochemistry is analysed on the Olympus AU400 according to the APF Clinical Chemistry SOP (-date-). Animals are bled at 10 weeks based on the previously scheduled bleed list. The Haematology bleed is a retro-orbital bleed of 200?l into EDTA-coated tubes (Sarstedt Microvette 200 K3E; Ref 20.1288). On receipt of samples, 100?l is removed and diluted with FACS Wash for haematology analysis, 5-10?l is taken for RBC FACS and the remaining volume is lysed for Lymphocyte FACS. Haematology is analysed on the Siemans Advia 2120 according to the APF Haematology SOP (-date-). The standard CBC-Differential analysis is performed. RBC FACS is analysed on the BD LSRII according to the APF RBC FACS SOP.