Next generation cloning

Recent advances in synthetic biology have provided tools to efficiently construct complex DNA molecules which are an important part of many molecular biology and biotechnology projects. The planning of such constructs has traditionally been done manually using a DNA sequence editor which becomes error-prone as scale and complexity of the construction increase.

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Traditional Map based Cloning approaches, used for the identification of desirable alleles, are extremely labour intensive and years can elapse between the mutagenesis and the detection of the polymorphism.

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Physical maps created from large insert DNA libraries, typically cloned in BAC vector, are valuable resources for map-based cloning and de novo genome sequencing. The maps are most useful if contigs of overlapping DNA clones are anchored to chromosome(s), and ordered along them using molecular markers.

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Plants are always under the radar of the pathogen, with their different means of infecting them. And conventional methods of managing plant diseases are showing transient effectiveness, as they never last long. Introduction of next generation sequencing, has provided with whole lot of information regarding host and pathogen. It analyses a large number of pathogen isolates and host varieties rapidly at lower cost. In this host pathogen dynamics, effector molecules play a very crucial role.

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MicroRNAs (miRNAs) are small, endogenously expressed, and nonprotein coding RNAs that regulate gene expression via post-transcriptional inhibition and cleavage. To date, several plant miRNAs have been identified via direct cloning, high-throughput sequencing, and bioinformatics analyses.

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There have been great strides in using positional cloning to successfully identify the underlying basis of many Mendelian disorders of children. The severity of even these disorders, however, is greatly affected by genetic modifiers and gene- environment interactions. The molecular pathogenesis of many more complex disorders remains unexplained due to the often complex genetic, epigenetic and environmental interactions involved in their etiology. However, there is reason for optimism.

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