Six PCR tips for quick mouse genotyping
Here are six PCR tips for quick mouse genotyping. 1. Skip genomic DNA extraction. Use a direct PCR master mix which allows you to add tissue directly to the PCR reaction, without the need for genomic DNA purification. 2. Optimize PCR annealing, faster. Use a direct PCR master mix that enables a universal annealing temperature of 60°C. 3. Handle fewer reagents tubes. Use a master mix format that is premixed with necessary PCR components. Only your sample and primers need to be added. 4. Finish PCR in minutes, not hours. Choose a polymerase that can synthesize DNA at 15–20 sec/kb for fast cycling. 5. Load PCR reactions directly onto gels. Use a direct PCR master mix containing electrophoresis dyes for direct gel loading. Skip addition of the dyes to the reactions for electrophoresis. 6. Complete electrophoresis in 15 min or less. Use bufferless precast gels and an electrophoresis system integrated with a camera hood. Just load, run, and analyze your PCR samples.