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Short and effective

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Cloning Vector : All the vectors used for propagation of DNA insert in a suitable host are known as cloning vector. Expression Vector : When a vector is designed for the expression of genes i.e., production of the protein specified by the concerned gene, it is termed as expression vector. As a rule expression vector must contain regulator sequences such as promoter, operator and ribosomal binding sites etc.

What is PCR? Polymerase chain reaction (PCR) is a thermodynamic as well as enzymatic process of production of billions copies of DNA from a single segment of DNA (µg). Kary Mullis in 1985 invented PCR which revolutionized the field of molecular biology. Kary Mullis demonstrated that oligonucleotide primers could be used to amplify specific segment of genomic DNA or cDNA. Mullis was awarded with Noble Prize in chemistry in 1993 for his contribution in development of PCR. Requirement of PCR Template DNA which is to be amplified. Nucleotide primers of about 20 bases long (Forward and Reverse both). Free nucleotides called ddNTPs (dATP, dGTP, dTTP and dCTP). A heat stable DNA polymerase e.g., Taq (Thermus aquaticus) polymerase, Pfu (Pyrococcus furiosus), Vent (Thermococus litoralis). Note : Pfu and Vent polymerase are more efficient than Taq polymerase. Application of PCR Gene fragment amplification as a quick alternative to cloning. DNA fragment modification Sensitive detection of harmful microbes, followed by accurate genotyping if desired. Archaeological specimen DNA analysis. DNA Fingerprinting and Marker assisted Selection Detection of transgene

What are plasmids? Plasmids are double stranded, circular, self replicating extra chromosomal DNA molecule, of bacterial cell. They have independent replication and transmission capacity. Exist either independent or may become integrated into the bacterial chromosome. DNA insert capacity 8-10 kb pBR 322 Developed by Bolivar and Rodriguez 1977 Size 4363 bp It has conjugation ability. Produces 15-30 copies / cell. Origin from ColE1 plasmid Two selectable marker tet & amp 20 unique recognition site 6 within tetR and 3 within ampR 2 within promoter region DNA insert integrated either in tet or amp

what are vector? Vectors are the carriers of genes which carry the genes from one organism to another organism and replicate it into many copies. Vector is a recombinant DNA molecule that is capable of independent replication when introduced into a suitable organism. Vector acts as a vehicle that transports the gene into a host cell (usually, bacterium). Ex. Plasmid, cosmid, phagemid, YAC and BAC etc.

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