PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation

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PLASMID ISOLATION AND ANALYSIS Part IIIPlasmid
Isolation
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Plasmid IsolationStep 1
Place a QIAprep spin column in the 2 ml
collection tube.
QIAprep spin column
collection tube
PLASMID ISOLATION AND ANALYSIS Part III
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Plasmid IsolationStep 2
Use the small needle pipette to carefully remove
all of the clear lysate/supernate from the L
tube.
PLASMID ISOLATION AND ANALYSIS Part III
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Plasmid IsolationStep 3
Add the clear lysate/supernate from tube L to
the QIAprep column.
PLASMID ISOLATION AND ANALYSIS Part III
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Plasmid IsolationStep 4
Centrifuge for 1 minute and discard the flow
through.
PLASMID ISOLATION AND ANALYSIS Part III
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Plasmid IsolationStep 5
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 AND ANALYSIS Part III
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Plasmid IsolationStep 6
Discard the flow through, and centrifuge for an
additional 1 minute to remove residual wash
buffer.
PLASMID ISOLATION AND ANALYSIS Part III
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Plasmid IsolationStep 7
Transfer the QIAprep spin column to the 1.5 ml
tube marked M in Part II, step 6.
QIAprep spin column
PLASMID ISOLATION AND ANALYSIS Part III
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Plasmid IsolationStep 8
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 AND ANALYSIS Part III
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Plasmid IsolationStep 9
Let stand for 1 minute and centrifuge for 1
minute to collect the DNA.
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Plasmid IsolationStep 10
Label the M tube with your groups name, seal
the tube with its original cap, and place in
freezer (-20oC) until you are ready to analyze
the extracted DNA in Part IV.
PLASMID ISOLATION AND ANALYSIS Part III