PCR was invented by Kary Mullis in 1983 and won Nobel Prize in 1993. PCR amplifies DNA millions to billions of times in temperature cycles. Requires primers and thermostable DNA polymerase for DNA synthesis
Manual covers DNA isolation, purification, and quantification methods. Provides protocols for gel electrophoresis and Southern blotting. Discusses plasmid vectors and transformation methods. Covers high-capacity vectors like Bacterial Artificial Chromosomes. Explains RNA extraction and analysis techniques
Sequence quality must be checked before primer selection. Repbase database helps identify vector contaminants and repeats. Low-quality bases should be converted to N's or excluded. Primer peaks often occur at sequence ends
NAATs detect small amounts of DNA or RNA in test samples. Tests use PCR or other methods to amplify pathogens. No living sample needed, quick testing possible
PCR copies DNA regions using Taq Polymerase and simple reagents. Taq polymerase withstands multiple heating/cooling cycles unlike other species. Requires template DNA, Taq polymerase, dNTPs, and primers. Creates thousands of identical copies from single template
Tool searches for specific primers against PCR templates using Primer3 and BLAST. Checks both forward-reverse and reverse-forward primer pairs. Can be set to automatically or manually guide primer-blast searches