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Phage display cycle. 1) fusion proteins for a viral coat protein + the gene to be evolved (typically an antibody fragment) are expressed in bacteriophage. 2) the library of phage are washed over an immobilised target. 3) the remaining high-affinity binders are used to infect bacteria. 4) the genes encoding the high-affinity binders are isolated.
Construction of a genomic library involves creating many recombinant DNA molecules. An organism's genomic DNA is extracted and then digested with a restriction enzyme.For organisms with very small genomes (~10 kb), the digested fragments can be separated by gel electrophoresis.
Construction of pre-made phage display libraries. Pre-made phage display library [ 10 ] is a laboratory technique for the study of protein–protein, protein–peptide, and protein–DNA interactions that uses bacteriophages (viruses that infect bacteria) to connect proteins with the genetic information that encodes them.
After the construction of recombinant lambda or cosmid libraries the total DNA is transferred into an appropriate E. coli host via a technique called in vitro packaging. The necessary packaging extracts are derived from E. coli cI857 lysogens (red- gam- Sam and Dam (head assembly) and Eam (tail assembly) respectively).
Later improvements of antibody phage display technology enables the display of millions of different antibody fragments on the surface of filamentous phage (better known as antibody phage library) and subsequent selection of highly specific recombinant antibodies to any given target. This technology is widely exploited in pharmaceutical ...
A P1-derived artificial chromosome, or PAC, is a DNA construct derived from the DNA of P1 bacteriophages and Bacterial artificial chromosome.It can carry large amounts (about 100–300 kilobases) of other sequences for a variety of bioengineering purposes in bacteria.
A genomic library is a set of clones that together represents the entire genome of a given organism. The number of clones that constitute a genomic library depends on (1) the size of the genome in question and (2) the insert size tolerated by the particular cloning vector system. For most practical purposes, the tissue source of the genomic DNA ...
YoctoReactor library size. yR library size is a function of the number of different functionalized oligos used in each position and the number of positions in the DNA junction. The yR design approach provides an unvarying reaction site with regard to both (a) distance between reactants and (b) sequence environment surrounding the reaction site.