How can you determine a rat's blood type? - briefly
Collect a small blood sample from the rat (commonly via tail‑vein puncture) and apply it to a rodent‑specific blood‑typing kit that uses antigen‑specific antisera to observe agglutination. The pattern of agglutination indicates the rat’s blood group.
How can you determine a rat's blood type? - in detail
Determining a rat’s blood group requires a controlled collection of a small blood sample, appropriate anticoagulation, and a reliable assay that matches the rodent’s antigenic system.
First, obtain the specimen under aseptic conditions. Anesthetize the animal with isoflurane or a ketamine‑xylazine mixture. Preferred collection sites are the lateral tail vein, saphenous vein, or retro‑orbital sinus. Use a 27‑ or 30‑gauge needle to draw 0.1–0.2 mL of blood into a microtube pre‑filled with 3–5 µL of EDTA or heparin to prevent clotting. Store the sample on ice and process within two hours.
Second, select an assay compatible with the rat blood‑group system (RT1). Two widely accepted methods are:
- Serologic agglutination – Mix the plasma or serum with a panel of rat‑specific anti‑RT1 antibodies on a glass slide or in a microtiter plate. Observe for clumping within 1–2 minutes. Positive agglutination indicates the presence of the corresponding antigen. Include positive and negative controls to validate the reaction.
- Polymerase chain reaction (PCR) genotyping – Extract genomic DNA from the same sample using a commercial kit. Amplify the RT1 locus with primers targeting known alleles (e.g., RT1.A, RT1.B, RT1.C). Separate products by agarose gel electrophoresis or perform real‑time PCR with allele‑specific probes. The pattern of amplification identifies the genotype.
Third, verify results by cross‑matching. Combine a small volume of the test plasma with red cells from a donor of known type; lack of agglutination confirms compatibility. Repeat the serologic test with a second antibody panel to rule out false‑positive reactions.
Finally, record the blood type in a laboratory notebook, noting the animal’s identifier, collection date, assay used, and any observed discrepancies. Store remaining blood at –80 °C for future molecular analysis if needed.