DNA Structure and Analysis
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Transcript of DNA Structure and Analysis
DNA Structure and Analysis
Activity 4.1: Restriction Site Prediction Using NEBcutter
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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
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Skills to Master
Use NEBcutter software to predict restriction enzyme sites
Record detailed software instructions into a laboratory notebook
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Student Workstation Materials
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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
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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