Changes

DNA Preparation from Tail Clip

23 bytes removed, 17:58, 5 June 2009
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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.