DNA Preparation from Tail Clip: Difference between revisions
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Use Qiagen DNEasy kit for Blood and Tissue Samples | Use Qiagen DNEasy kit for Blood and Tissue Samples. For a crude preparation, often good enough for PCR genotyping see [[Preparation of Tail Samples (for Genotyping)]] | ||
==Protocol== | ==Protocol== | ||
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#Place DNeasy Mini spin column in new collection tube and add 500 ul Buffer AW2. Centrifuge for 3 minutes at 14000rpm. Discard flow-through and collection tube. | #Place DNeasy Mini spin column in new collection tube and add 500 ul Buffer AW2. Centrifuge for 3 minutes at 14000rpm. Discard flow-through and collection tube. | ||
#Place DNeasy Mini spin column in clean 1.5 microcentrifuge tube and pipet 200ul Buffer AE directly onto the DNeasy membrance. Incubate at room tempature for 1 minute and then centrifuge for 1 minute at 8000rpm to elute. | #Place DNeasy Mini spin column in clean 1.5 microcentrifuge tube and pipet 200ul Buffer AE directly onto the DNeasy membrance. Incubate at room tempature for 1 minute and then centrifuge for 1 minute at 8000rpm to elute. | ||
[[Category: Mouse Work]] | |||
[[Category: Molecular Biology]] | |||
[[Category: DNA]] | |||
[[Category: Genotyping]] | |||
Latest revision as of 19:23, 24 January 2026
Use Qiagen DNEasy kit for Blood and Tissue Samples. For a crude preparation, often good enough for PCR genotyping see Preparation of Tail Samples (for Genotyping)
Protocol
- Cut .5cm lengths of tail into a 1.5 ml microcentrifuge tube. Add 180ul Buffer ATL.
- Add 20ul proteinase K. Mix by vortexing and incubate at 56C until tissue is completely lysed. (Overnight)
- Vortex for 15s. Add 200ul Buffer Al to sample and vortex. Then add 200 ul ethanol (96-100%) and vortex.
- Pipet the mixture from step 3 into DNeasy Mini spin column. Centrifuge at 8000rpm for 1 minute. Discard flow-through and collection tube.
- Place DNeasy Mini spin column in new collection tube and add 500 ul Buffer AW1. Centrifuge for 1 minute at 8000rpm. Discard flow-through and collection tube.
- Place DNeasy Mini spin column in new collection tube and add 500 ul Buffer AW2. Centrifuge for 3 minutes at 14000rpm. Discard flow-through and collection tube.
- Place DNeasy Mini spin column in clean 1.5 microcentrifuge tube and pipet 200ul Buffer AE directly onto the DNeasy membrance. Incubate at room tempature for 1 minute and then centrifuge for 1 minute at 8000rpm to elute.