12/12/2016
BASIC OVERVIEW OF BIOTECHNOLOGYThe term "Bio" refers to life or living organism and"technology" is related to mechanics and computing.So biotechnology is the use of science and techniques to make products and modified processes. Biotechnology is based on the idea that all living organism has something in common. The common thing among all living organism is the DNA. So what makes one living organism different from other is the order of nitrogen bases.GENETIC ENGINEERING--------------------------------If all the living organism is made up of DNA, then it should be easy to mix and match the DNA from different organism. This is called genetic engineering.RESTRICTION ENZYMES---------------------------------In order to mix and match the genes, we need to cutthe DNA using DNA scissors. In biotechnology thereare specific types of molecules that can act as scissors. They are called restriction enzymes. Enzymes are molecules that speed up or catalyses chemical reactions. Restriction enzyme is a molecule that speeds up a reaction that cuts DNA.DNA LIGASE-----------------DNA ligase is a specific type of enzyme that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond.RECOMBINANT DNA----------------------------Recombinant DNA, or rDNA, is the term used to describe the combination of two DNA strands that are constructed artificially from different organism. Genetic scientists can do this to create unique DNA strand for different purposes, using several types of techniques. Like naturally occurring DNA, recombinant DNA has the ability to produce recombinant proteins.TRANSGENIC ORGANISM----------------------------------The organism that gets the DNA from other organism is called transgenic organism. Organisms that have altered genomes are known as transgenic.CLONE----------In biotechnology, clone is any living organism with genetic material that is identical to that of the parent organism from which it was created.GEL ELECTROPOROSIS---------------------------------Gel electrophoresisis a laboratory method used to separate mixtures of DNA, RNA, or proteins according to molecular size. In gel electrophoresis, the molecules to be separated are pushed by an electrical field through a gel(agarose gel) that contains small pores.RIBONUCLEASE----------------------Ribonuclease or RNase is an RNA digesting enzymethat catalyzes the degradation of RNA into smaller components.TAQ DNA POLYMERSASE----------------------------------Taq DNA polymerase is isolated from Thermus aquaticus YT1. Taq DNA polymerase is heat stable and will synthesize DNA at elevated temperatures from single stranded templates in the presence of primer.LINKERS------------A linker is a short piece of synthesized double stranded DNA that has unique site for one or more restriction enzymes. It is a blunt ended molecule and it yields cohesive ends while cutting it with a suitable restriction enzyme. It is used to join blunt ended DNA's.ADAPTORS---------------An adapter is a short double stranded DNA that has one blunt end and one cohesive end. The cohesive end is the same as the cut site produced by a particular restriction enzyme.PLASMIDS--------------Plasmids are circular double stranded extra chromosomal DNA's present in bacterial cells. Theyreplicate independently due to the presence of an origin of replication. Plasmids are used in the laboratory manipulation of genes.COSMIDS-------------A cosmid is a type of hybrid plasmid that contains a Lambda phage cos(cohesive end sites) sequence.Cosmids = cos sites plasmid.YEAST ARTIFICIAL CHROMOSOMES------------------------------------------------Yeast artificial chromosomes (YACs) are genetically engineered chromosomes derived from the DNA of the yeast, Saccharomyces cerevisiae, which is then ligated into a bacterial plasmid.BACTERIAL ARTIFICIAL CHROMOSOMES-------------------------------------------------------Bacterial artificial chromosome(BAC)is the synthetic plasmid that behaves as an additional genetic material in bacteria. The BAC exists as a single copy in bacterial cell and is inherited from parent to daughter cells during cell division. BAC was first developed in 1990 to clone large DNA's in E.coli.POLYMERASE CHAIN REACTION--------------------------------------------Polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy 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.MICROINJECTION------------------------Microinjection refers to the injection of DNA's or cell organelles directly into cells using a injection needle. By this method RNA's, DNA's, proteins or cell organelles are injected into animal cells, eggs, zygotes and pant protoplasts.CHROMOSOMAL WALKING------------------------------------Chromosomal walking is a technique used to isolateunknown sequences found next to a known gene.CHROMOSOMAL JUMPING------------------------------------Chromosomal jumping is used for finding widely spaced gene loci by cloning long DNA segments. It helps for fast reading of loci along the length of the chromosomal DNA. This is also know as gene jumping.SOUTHERN BLOTTING------------------------------Southern blotting is used to detect the presence of aparticular DNA fragment in a sample. The DNA detected can be a single gene, or it can be a part of a large piece of DNA. This technique was developedby E.M Southern in 1975.NORTHERN BLOTTING-------------------------------Northern blotting is used to detect the presence of mRNA in a sample. It was developed by Alwine and his colleagues in 1979.WESTERN BLOTTING-----------------------------Western blot is often used in research to separate and identify proteins. In this technique a mixture of proteins is separated based on molecular weight, and thus by type, through gel electrophoresisCAT GENE--------------The enzyme chloramphenicolacetyl transferase(CAT) is encoded by CAT gene. It provides resistance to E.Coli against Chloramphenicol.ENZYME LINKED IMMUNO SORBENT ASSAY-----------------------------------------------------------Enzyme-linked immunosorbent assay (ELISA), also known as an enzyme immunoassay (EIA), is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample.DNA SEQUENCING-------------------------DNA sequencing is the process of determining the precise order of nucleotides within a DNA molecule.It includes any method or technology that is used to determine the order of the four bases adenine, guanine, cytosine, and thymine in a strand of DNA.SATELLITE DNA---------------------Satellite DNA is a non coding region of eukaryotic cells having tandem repeats of a sequence.Tandemrepeats occur in DNA when a pattern of one or morenucleotides is repeated and the repetitions are directly adjacent to each other. Satellite DNA is alsocalled mini-satellite.It is rich in GC pairs and 9-40bpin size.PROBE----------In molecular biology, a hybridization probe is a fragment of DNA or RNA of variable length (usually 100–1000 bases long) which can be radioactively labeled. It can then be used in DNA or RNA samplesto detect the presence of nucleotide sequences (theDNA target) that are complementary to the sequence in the probe.ZOO BLOT--------------Zoo blot is obtaining the images of the DNA of animal on x-ray film.GARDEN BLOT--------------------Garden blot is obtaining the images of the DNA of plant on x-ray film.RANDOM AMPLIFIED POLYMORPHIC DNA's(RAPD's)----------------------------------------------------------------------RAPDs are DNA fragments amplified by PCR using short synthetic primers (generally 10 bp) of random sequence. These oligonucleotides serve as both forward and reverse primer, and are usually able to amplify fragments from 1-10 genomic sites simultaneously.VARIABLE NUMBER OF TANDEM REPEATS(VNTRs)--------------------------------------------------------------------DNA of eukaryotic organisms has several repeating units of short sequences called tandem repeats. The variation in the number of tandem repeats between two or more individuals is called variable number of tandem
