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Edvotek 112 Analysis of Eco RI Cleavage Patterns of Lambda DNA

Biotechnology - Restriction Enzyme Analysis of DNA - Edvotek 112/AP09-112

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This Edvotek experiment introduces the use of restriction enzymes as a tool to digest DNA at specific nucleotide sequences. Bacteriophage lambda DNA has a linear structure and 6 Eco RI recognition sites. Separation by agarose gel electrophoresis of an Eco RI digest of lambda DNA will yield 6 bands (5 distinct bands, two are very close in size) corresponding to the DNA fragments.

• Introduce restriction enzymes and their ability to digest DNA at specific nucleotide sequences
• Understand how electrophoresis separates differently-sized molecules
• Perform agarose gel electrophoresis
• Load, run, analyze, compare and size lambda DNA cut with HindIII, lambda DNA cut with EcoRI, and uncut lambda DNA

Kit includes: Instructions, DNA samples (packaged as either Pre-aliquoted QuickStrip™ connected tubes or as individual 1.5 ml (or 0.5 ml) microcentrifuge tubes), UltraSpec-Agarose™, Electrophoresis Buffer (50X), Practice Gel Loading Solution, FlashBlue™ DNA Stain.

All you need: DNA Electrophoresis, Micropipettes: 5-50 µl (Optional), White Light Box, & Microwave or Hot Plate.

Time required: 60 minutes.

For 8 Gels.