If you do bench work in a life science lab, you’ve likely delved into molecular biology techniques (PCR, cloning, site-directed mutagenesis). Ask any scientist; they have a love/hate relationship with it. Love it when it works, hate it when it doesn’t.
Most biologists use PCR for two reasons:
1) Cloning: Amplifying an insert piece of DNA to subsequently cut with restriction enzymes and paste into a plasmid.
2) Quick changes (site-directed mutagenesis): Introducing a point mutation, small insert, or deletion into an intact plasmid
As a fifth-year PhD candidate, I’ve done my fair share of cloning and site-directed mutagenesis. Not to toot my own horn, but my success rate is pretty high. Why, you may ask? Here’s why:
1. Primer Design Hacks
2. PCR Hacks
So there you have it! My tips for increasing your PCR success rate. Give them a try!!