Viagen Biotech Inc
Direct PCR (Tail)
102-T
Aim of the present study was to do genotyping of male albino wistar rats for further experimental uses.
Genotyping of rats
Rats tail, 1.5 ml centrifuge tube, Direct PCR (Tail) reagent, Proteinase K, Centrifuge, PCR mastermix, genotyping primers, nuclease free water, PCR machine, agarose gel electrophoresis
Rats of 5 weeks age maintained on standard management conditions were anesthetized with isofurane, a small piece of tail (2mm) was cleaned with 10% ethanol and sampled. Cut piece of tail were submerged in 100 micL of direct PCT (Tail) reagent mixed with proteinase K and incubated for overnight at 54 degree Celsius. After incubation, proteinase K was denatured by incubating samples at 85 degree celsius for 45 minutes. Digested samples were centrifuged at 2000 xg for 5 min and supernatant were collected in another fresh centrifuge tube. Two micL of supernatant were mixed with PCR master mix along with genotyping primers and PCR was performed. PCR product was run in 2% agarose gel and visualized in UV light.
No
A good quality PCR amplification products were visualized in UV light. The Direct PCR (Tail) does not interfere with DNA polymerase activity as very neat and clean bands of PCR products were visualized in agarose gel electrophoresis.
Fast and quick method to perform genotyping by omitting DNA isolation step
Best quality Direct PCR reagent.