Techniques In Plant Biotechnology By S. Datta, P.G. Patil, K.R. Soren, Alok Das, S.K. Chaturvedi, N. Nadarajan

Techniques In Plant Biotechnology By S. Datta, P.G. Patil, K.R. Soren, Alok Das, S.K. Chaturvedi, N. Nadarajan
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Techniques In Plant Biotechnology By S. Datta, P.G. Patil, K.R. Soren, Alok Das, S.K. Chaturvedi, N. Nadarajan

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Publisher: Agrotech Publications
ISBN: 9788183212793
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PREFACE The need of working biotechnological protocols is must for research workers, graduate and post graduate students of plant biotechnology. The present manual is an attempt in this direction to meet these requirements. The manual covers all aspects of plant molecular biology like molecular marker techniques, functional genomics, recombinant DNA technology and transgenics. In this manual all the working protocols are provided with their underlying basic principles that can be easily followed by researchers and students. In this manual, the basic microbiological techniques for isolation of single bacterial colony to quantification by streak plate and turbidimetric has been described in the first three experiments. The next experiments mainly deals with molecular marker techniques beginning with genomic DNA isolation and quantification followed by various PCR and restriction digestion based techniques like random amplified polymorphic DNA RAPD , simple sequence repeats SSRs and amplified fragment length polymorphism AFLP and finally scoring and interpretation of the molecular data for diversity studies. We then moved towards recent genomics approaches for construction of expression libraries right from isolation and quantification of total RNA from leaf tissues to construction of cDNA libraries. The next set of experiments deals with basics of recombinant DNA techno logy rDNA like cloning, transformation and characterization of recombinant clones from E.coli. The basic and applied aspects of plant tissue culture beginning with preparation of stock solutions and their storage, culture and maintenance of Agrobacterium strains, direct and indirect methods of gene transfer in pulses and quick histochemical analysis are dealt with in next few experiments. Finally, valuable resources in bioinformatics available for various applications are also being highlighted. We have also provided readymade recipes of commonly used buffers and solutions to be used in the various experiments. For quick reference of the readers, a comprehensive glossary covering all the topics are provided at the end of the manual. Over all these protocols can be used in isolation and characterization of novel
iv genes, germplasm characterization, mapping of important biotic and abiotic stress resistance genes and direct and indirect genes transfer methods in developing transgenic plants. Though we have used pulses as experimental material throughout the experiments, we hope these protocols will work equally well for other plant species. We wish to thank many people who have made efforts in bringing out this manual in a lucid format. Special thanks to all our students and staffs for their valuable inputs at different stages of preparation of this manual. We hope that these protocols will make considerable impact among working