Techniques Used in Molecular Probing, Gel Electrophoresis, Genomic DNA, Plant Animal Species, Molecular Probes, High Voltage Electric Current

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Chimeric DNA Molecular Probes and Gene Libraries Chimeric DNA Molecular Probes and Gene Libraries Introduction Restriction Enzymes for Cloning Technique of Restriction Mapping Restriction Clevage and Gel Electrophoresis Construction of a Restriction Map Use of Partial Digests end Labelling and Hybridization in Restriction Mapping Construction of Chimeric DNA Cloning of Plasmid Vectors Directional Cloning of DNA Fragments with Heterologus Protruding Termini Adding Poly dA at 3' Ends of DNA Clone Blunt End Ligation by T4 DNA Ligase Cloning in Phage λ and Cosmid Vectors Cloning in phage λ Vectors Cloning in Cosmid Vectors Hosts for Cloning Vectors Bacteria as Hosts Eukaryotes as Hosts Molecular Probes Preparation of Probes Genomic DNA Probes

Home >> Biotechnology and Genomics >> Chimeric DNA Molecular Probes and Gene Libraries >> Techniques Used in Molecular Probing

Your Ad Here	Techniques used in Molecular Probing Separation of DNA fragments using agarose or polyacrylamide gel electrophoresis (PAGE). When genomic DNA, extracted from any tissue of a plant or animal species is digested with a restriction enzyme it is cleaved into segments. The segments of different sizes can be separated through gel electrophore is before a molecular probe is used to detec1 the segments which have sequences similar those in the probe. Gel electrophoresis involves movement of fragments or molecules under a high voltage electric current. The mixture of DNA fragments is loaded in a well created on one edge of the gel.
The gel may be a cylinder or a slab (usually a slab for cloning experiment), about 10 cm long and 0.5 cm thick. The rate of movement of fragments is inversely correlated with the size of fragments or molecules, so that heavier fragments will remain closer to the site of loading and the lighter fragments will move away Fragments of different sizes will appear as bands on the gel and can be examined or isolated for further study. More often agarose gels and can be examined or isolated for further study. More often agarose gels are used, but for separation of fragments differing by few base pairs, polyacrylamide gels are used Polyacrylamide gels are more commonly used for DNA sequencing experiments.
A gel with electrophoresed DNA in several lanes Separation of large DNA molecules (whole chromosomes) using PFGE The technique of agarose/polyacrylamide gel electrophoresis is used for separation of DNA molecules of different sizes. However, DNA molecules of large size could not be handled in this technique. A new technique called pulsed field gel electrophoresis (PFGE) has been used, for separation of large sized DNA moelcules, sometimes representing whole chromosomes. Using this technique, separation of DNA molecules belonging to each of the 16 individual chromosomes of yeast has become possible.	Your Ad Here
A gel with 12 bands representing 15 chromosomes of yeast as resolved by PFGE In this technique, short pulses of electricity are used in two different directions and DNA is embedded and used in the form of agarose plugs to avoid fragmentation of large DNA molecules. Using the technique of PFGE and a more refined technique CHEFE (countour clamped homogeneous electric field electrophoresis), genomes of several fungi could .be resolved into chromosomal bands and used for mapping of DNA sequences on specific chromosomes.

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c DNA Probes Synthetic Oligonucleotides as Probes RNA Probes or Riboprobes Labelling of Probes Radiolablled Probes Non Radioactive Probes Biotin Labelled Probes Digoxigenin Labelled Probes Alternatives to Biotin and Digoxigenin Labelling Amplification of DNA Probe Signals Techniques Used in Molecular Probing Southern Nothern and Western Blotting Southern Blotting Northern Blotting Western Blotting Dot Blots and Slot Blots Applications of Molecular Probes Construction and Screening of Genomic and cDNA Libraries Genomic Library by Shotgun Experiment cDNA Library from mRNAs Colony or Plaque Hybridization for Screening of Libraries Chromosome Walking and Characterization of Chromosome Segments Chromosome Jumping or Hopping Libraries BAC Libraries and Assembly of BACs into Contigs

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