Description:

Genomic DNA libraries are almost always screened by hybridization using a radioactive nucleic acid probe. Since this approach is essentially independent of a particular vector or type of target DNA, the main problem faced when considering creation of a genomic DNA library is simply generating a large enough number of recombinant DNA clones. The basic strategies used to address this problem have included both minimizing the number of clones necessary by incorporating large fragments of genomic DNA, and maximizing cloning efficiency by using vectors based on bacteriophage. This unit discusses the appropriate numerical considerations for both ordinary genomic DNA libraries and sub genomic DNA libraries, and then describes a limited number of appropriate vectors.

Genomic Deoxyribonucleic Acid (DNA) can be prepared from any source by three steps: cell lysis, deproteinisation and recovery of DNA. The basic protocol needs to be adapted to the demands of the application, the number of samples to be processed, the requested yield, purity and molecular weight of the DNA and the amount and history of the source. Traditional protocols based on the lysis with sodium dodecyl sulphate/proteinase K, extraction with phenol/chloroform/isoamyl alcohol, purification with ethanol and storage in tris/ethylenediamine tetraacetic acid buffer are recommended for the purification of long-term stable high-molecular weight genomic DNA from freshly obtained specimens. Intact chromosomal DNA is recovered from agarose-embedded cells. Automated extraction methods are preferred for forensic applications and high-throughput processing for bio banking.  The technically challenging recovery of ancient DNA needs to be optimised on a case-to-case basis, because ancient DNA is damaged and chemically modified and contains large amounts of polymerase chain reaction inhibitors.

A simple, practical method to watermark short trademarks or signatures into genomic DNA is introduced. Since the marking method is biologically innocuous, it can be applied to all commercialized bacteria to help establish brand names for the engineered strains and to resolve legal disputes regarding gene-related patents. The first such strain of Bacillus subtilis is engineered and is ready to be distributed. By virtue of the powerful technology developed in molecular biology, it is possible to isolate any DNA fragment in the genome of an organism and, after reverse transcription, any transcribed gene in the form of a complementary DNA. The isolation (cloning) procedure involves the insertion of the DNA fragment into a vector, capable of replication in a microorganism, which allows production of large quantities of the DNA fragment for physical or biological analysis. Upon determination of the location in the genome from which the particular DNA fragment was derived, that fragment acquires the property of a DNA marker. Such DNA markers are a prerequisite for physical and genetic mapping of the genome of the organism. DNA markers are also of importance for the diagnosis of genetic diseases. DNA markers can be divided into several different classes depending on the way in which the markers were selected among the fragments of genomic DNA. Examples of such classes are anonymous, micro- and minisatellites, Restriction Fragment Length Polymorphism (RFLP) markers, and NotI linking clones.

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With Regards

William 
Journal Coordinator
Global Journal of Research and Review