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DNA Structure and Analysis

DNA Structure and Analysis. Activity 4.1: Restriction Site Prediction Using NEBcutter . Activity 4.1: Restriction Site Prediction Using NEBcutter. Research Questions: How many times does a given restriction enzyme cut lambda DNA?

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DNA Structure and Analysis

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  1. DNA Structure and Analysis Activity 4.1: Restriction Site Prediction Using NEBcutter

  2. Activity 4.1: Restriction Site Prediction Using NEBcutter • Research Questions: • How many times does a given restriction enzyme cut lambda DNA? • When cutting lambda DNA with a given restriction enzyme, what pattern will the resulting DNA fragments make on a gel? • Objectives: • Use NEBcutter to predict where restriction enzymes will cut the bacteriophage lambda DNA sequence • Produce a virtual gel image of lambda DNA cut with a specific restriction enzyme

  3. Skills to Master • Use NEBcutter software to predict restriction enzyme sites • Record detailed software instructions into a laboratory notebook

  4. Student Workstation Materials

  5. Protocol Highlights/Tips • Go to the NEB website • tools.neb.com/NEBcutter2/ • Select Lambda under Standard sequences • Ensure that the radio button for Linear sequence is selected • Write detailed steps into notebook

  6. Summary • Make sure to: • Record all steps in your notebook • Lambda sequence is selected under standard sequences • Sequence is selected as linear • Print the fragments list and gel image to paste into your notebook • Adjust the gel type to 1% agarose and adjust the marker to Lambda-HindIII Digest • DNA type should be unmethylated

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