Molecular biology techniques: Polymerase chain reaction, Plasmid, Southern blot, Northern blot, Gene knockout, Western blot, Cell culture, DNA sequencing, DNA microarray, Immunoprecipitation, Transmission electron microscopy DNA sequencing

Source: Wikipedia. Pages: 92. Chapters: Polymerase chain reaction, Plasmid, Southern blot, Northern blot, Gene knockout, Western blot, Cell culture, DNA sequencing, DNA microarray, Immunoprecipitation, Transmission electron microscopy DNA sequencing, Methods to investigate protein¿protein interactions, Exome sequencing, ChIP-on-chip, Förster resonance energy transfer, Nucleic acid structure determination, Chromatin immunoprecipitation, Eastern blotting, ChIA-PET, COLD-PCR, Paired-end Tags, Combined bisulfite restriction analysis, Primer dimer, Kodecyte, DNA footprinting, Suspension array technology, PBR322, Allele-specific oligonucleotide, Chip-Sequencing, Streptamer, Oligomer restriction, Reverse transfection, Strep-tag, Selection and amplification binding assay, Knockout moss, Native PAGE, Isoelectric focusing, Helicase-dependent amplification, Subcloning, Plant transformation vector, Chemically defined medium, Magnet-assisted transfection, Branched DNA assay, TA cloning, Nuclease protection assay, Bio-Layer Interferometry, Southwestern blot, Synchronous culture, MassTag-PCR, Far-Eastern blotting, Ribosomal Intergenic Spacer analysis, Ligase chain reaction, Isopeptag, Far-western blotting, Gene knockin, Baby hamster kidney cell, Univec, VectorDB, Chromogenic in situ hybridization, Alanine scanning, Calcium chloride transformation, Fluorophore assisted carbohydrate electrophoresis, Reverse northern blot, Gel Doc, Racker unit, Promoter bashing, PBLU. Excerpt: The polymerase chain reaction (PCR) is a scientific technique in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. Developed in 1983 by Kary Mullis, PCR is now a common and often indispensable technique used in medical and biological research labs for a variety of applications. These include DNA cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; the diagnosis of hereditary diseases; the identification of genetic fingerprints (used in forensic sciences and paternity testing); and the detection and diagnosis of infectious diseases. In 1993, Mullis was awarded the Nobel Prize in Chemistry along with Michael Smith for his work on PCR. The method relies on thermal cycling, consisting of cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA. Primers (short DNA fragments) containing sequences complementary to the target region along with a DNA polymerase (after which the method is named) are key components to enable selective and repeated amplification. As PCR progresses, the DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the DNA template is exponentially amplified. PCR can be extensively modified to perform a wide array of genetic manipulations. Almost all PCR applications employ a heat-stable DNA polymerase, such as Taq polymerase, an enzyme originally isolated from the bacterium Thermus aquaticus. This DNA polymerase enzymatically assembles a new DNA strand from DNA building-blocks, the nucleotides, by using single-stranded DNA as a template and DNA oligonucleotides (also called DNA primers), which are required for initiation of DNA synthesis. The vast majority of PCR methods use thermal cycling, i.e., alternately heating and cooling the PCR sample to a defined series of temperature steps. These thermal cy