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文献和实验L; 3)IMAC fractions: 1 μL for Wnt3A immunoblot (or 10 μL for Coomassie-stained gel); 4) gel filtration fractions: 2 μL forWnt3A immunoblot (or 15 μL for Coomassie-stained gel); 5)heparin fractions: 1 μL for Wnt3A immunoblot or Coomassie-stained gel
Preparation of End-Labeled DNA Probes by Conventional Kinase for DNA Footprinting Analysis
Treatment 1. Add 20 μl of 10X CIP Buffer. 2. Add diluted CIP stock to the DNA such that the final concentration is approximately 0.2 Units CIP per μg DNA in a final volume of 200 μl (see Hint #4). 3. Incubate at 37°C
. At this point, you may want to collect a 2μl sample of your probe (from the 100 μl), and combine it with 10 μl 1X SDS gel loading buffer. You can then run this sample on a gel, and put film on the dried gel to see if the probe labelling step actually worked
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