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Biologie
Isolation of Genomic DNA from Mouse Tails
Published: July 29, 2007
doi:
10.3791/246
Tony Zangala
1
Department of Physiology and Biophysics
,
University of California, Irvine (UCI)
Protocol
Cut tail pieces (3mm) and mark ears.
Add 720
m
l STE and 30
m
l Proteinase K (10 mg/ml stock).
Incubate at 55
°
C on heating block and vortex every hour for 3 hours at top speed for 10 seconds.
Inactivate proteinase K at 70
°
C for 5 minutes.
Quench on ice for 5 minutes.
Centrifuge tail DNA for 10 minutes at full speed.
Decant into new tube containing 720
m
l Isopropanol.
Precipitate DNA by inverting the tube or vortexing.
Spin down DNA for 5 minutes at full speed.
Remove supernatant.
Wash pellet with 70% ethanol.
Spin down genomic DNA 5 minutes at full speed.
Remove supernatant.
Allow DNA to dry for 1-2 minutes.
Resuspend DNA in 100-200
m
l depending on size of pellet.
Place tube at 55
°
C for 1 hour to facilitate dissolution of DNA.
Materials
Material Name
Typ
Company
Catalogue Number
Comment
STE
Buffer
100 mm Tris; pH 8.5 / 5 mM EDTA / 0.2% SDS / 200 mM NaCl / in H2O
TE
Buffer
100 mM Tris; pH 5.0 / 1 mM EDTA / 10 mM NaCl
DOWNLOAD MATERIALS LIST
Tags
Isolation
Genomic DNA
Mouse Tails
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DOI
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Diesen Artikel zitieren
Zangala, T. Isolation of Genomic DNA from Mouse Tails.
J. Vis. Exp.
(6), e246, doi:10.3791/246 (2007).
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