A plasmid having a restriction site for Hind III enzyme within a kanamycin resistance gene was cleaved with Hind III and was re-ligated. It was used to transform E.coli (Escherichia coli) K12 cells. From the culture plate, kanamycin resistant colonies were selected and electrophoresis of the plasmid DNA from these colonies was carried out. Most of the colonies with plasmid produce a single band, which migrated at the same speed as the original intact plasmid. A slow band in agarose gel electrophoresis was obtained as a product of ligation. Introduction: Agarose gel electrophoresis is standard lab procedure for a DNA (deoxyribonucleic acid) separation on the basis of size (length in base pair). Agarose gel electrophoresis uses an electrical field to mobilize negatively charged DNA molecule in an agarose gel matrix toward the positively charged electrode. A DNA molecule, when digested with restriction enzymes (tools of recombinant DNA technology or RDT), produces the bands of different sizes when subjected to electrophoresis. This technique is used for diagnostic purposes to determine the genetic defects.

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Chapter 17, Problem 13PDQ

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A plasmid having a restriction site for Hind III enzyme within a kanamycin resistance gene was cleaved with Hind III and was re-ligated. It was used to transform E.coli (Escherichia coli) K12 cells. From the culture plate, kanamycin resistant colonies were selected and electrophoresis of the plasmid DNA from these colonies was carried out. Most of the colonies with plasmid produce a single band, which migrated at the same speed as the original intact plasmid. A slow band in agarose gel electrophoresis was obtained as a product of ligation.

Introduction:

Agarose gel electrophoresis is standard lab procedure for a DNA (deoxyribonucleic acid) separation on the basis of size (length in base pair). Agarose gel electrophoresis uses an electrical field to mobilize negatively charged DNA molecule in an agarose gel matrix toward the positively charged electrode. A DNA molecule, when digested with restriction enzymes (tools of recombinant DNA technology or RDT), produces the bands of different sizes when subjected to electrophoresis. This technique is used for diagnostic purposes to determine the genetic defects.

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