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==annealing with AluI oligo and digestion with ALuI(8/25/08)== 1. Add 1ul NEB buffer2, 1 uL of 100 uM RE-AluI-V6_2_Ida guide oligo in 5ul circularized probe from previous steps. 2. Denature at 95C in PCR machine for 10 mins. Turn off the machine and wait for 20 min. 3. Add 2uL of AluI restriction enzyme to reaction, incubate at 37C for about 2 hours. [[Image:8-25-08 after alu-1 digestion.jpg]] Note: the left most lane had some urea in the well while loading the gel, so not all the sample traveled down to the bottom of the well when starting the gel, thus might have affected the result of the gel. Theoretically, two lanes should look quite similar based on previous results.
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