Week 4_handout

Week 4_handout - Bi 1x Spring 2010 Week 4 Session 2 o...

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Bi 1x, Spring 2010 Week 4 Session 2 o Restriction enzyme digestion Restriction enzyme digestion Restriction enzymes are enzymes that bind to specific DNA sequences and cleave (“digest”) the DNA at or next to the binding site. You will become familiar with restriction enzymes during this week’s lab sessions on both bulk and single molecule scales. Note: This document describes bulk restriction digests only. Materials: Sterile, ultrapure water pZE21-lacZ plasmid DNA 10X NEB2 Buffer 10X EcoRI Buffer 10X BSA Solution HindIII digested lambda phage DNA EcoRI, HindIII, and KpnI restriction enzymes will be supplied by your TAs when you are ready to use them. Background: Most useful restriction enzymes recognize 4-8 base pair restriction sites. These sites are symmetric, inverted repeats called palindromes. Shown below are the restriction sites of the three enzymes you will be using today: EcoRI, HindIII, and KpnI. Notice how the 5’ to 3’ sequence is identical on the top and bottom strands.
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