Regarding DNA preparation, Blake had good results using 1 mL 10^9 PFU High Titer phage preparations. His DNA was suspended into 100 uL of water at the end of the procedure, and he would use 5 uL of this DNA for restriction digestion. We will take the same approach. Each 20 uL cut will contain:
2 uL buffer,
1 uL BSA,
1 uL Enzyme,
5 uL DNA.
The digestion will occur at 37°C for 2-3 hours, and will then immediately be loaded onto a gel. During the 2-3 hour gap, we will run 5 uL of each DNA sample in a different gel to determine whether there is any DNA or not, running the DNA for 5 min at 100 V. No DNA? The student should have time to prepare new DNA today so that digestions can be done tomorrow with that DNA...the reason for having 2 digestion days.