PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation

PLASMID ISOLATION AND ANALYSIS: Part III Place a QIAprep ® spin column in the 2 ml collection tube. collection tube QIAprep ® spin column Plasmid Isolation Step 1

PLASMID ISOLATION AND ANALYSIS: Part III Use the small needle pipette to carefully remove all of the clear lysate/supernate from the “L” tube. Plasmid Isolation Step 2

PLASMID ISOLATION AND ANALYSIS: Part III Add the clear lysate/supernate from tube “L” to the QIAprep ® column. Plasmid Isolation Step 3

PLASMID ISOLATION AND ANALYSIS: Part III Centrifuge for 1 minute and discard the flow through. Plasmid Isolation Step 4

PLASMID ISOLATION AND ANALYSIS: Part III Using a small needle pipette, add the contents of the tube marked “Wash Buffer” (PE) to the QIAprep ® spin column and centrifuge for 1 minute. Plasmid Isolation Step 5

PLASMID ISOLATION AND ANALYSIS: Part III Discard the flow through, and centrifuge for an additional 1 minute to remove residual wash buffer. Plasmid Isolation Step 6

PLASMID ISOLATION AND ANALYSIS: Part III Transfer the QIAprep ® spin column to the 1.5 ml tube marked “M” in Part II, step 6. QIAprep ® spin column Plasmid Isolation Step 7

PLASMID ISOLATION AND ANALYSIS: Part III Use a small needle pipette to elude the DNA by adding the contents of the tube marked sterile water (SW) to the QIAprep ® spin column. Plasmid Isolation Step 8

PLASMID ISOLATION AND ANALYSIS: Part III Let stand for 1 minute and centrifuge for 1 minute to collect the DNA. Plasmid Isolation Step 9

PLASMID ISOLATION AND ANALYSIS: Part III Label the “M” tube with your group’s name, seal the tube with its original cap, and place in freezer (-20 o C) until you are ready to analyze the extracted DNA in Part IV. Plasmid Isolation Step 10