DNA Preparation from Tail Clip: Difference between revisions
Created page with 'Use Qiagen DNEasy kit for Blood and Tissue Samples' |
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Use Qiagen DNEasy kit for Blood and Tissue Samples | Use Qiagen DNEasy kit for Blood and Tissue Samples | ||
[edit] Protocol | |||
1. Cut .5cm lengths of tail into a 1.5 ml microcentrifuge tube. Add 180ul Buffer ATL. | |||
2. Add 20ul proteinase K. Mix by vortexing and incubate at 56C until tissue is completely lysed. (Overnight) | |||
3. Vortex for 15s. Add 200ul Buffer Al to sample and vortex. Then add 200 ul ethanol (96-100%) and vortex. | |||
4.Pipet the mixture from step 3 into DNeasy Mini spin column. Centrifuge at 8000rpm for 1 minute. Discard flow-through and collection tube. | |||
5. 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. | |||
6. 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. | |||
7. 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. | |||