NOTE: Each sequence will require two samples—one with the upstream primer and one with the downstream primer
Label 1.5 mL microcentrifuge tubes
Add 1 μL of the appropriate primer to each tube
Add DNA from a cleaned upPCR according to the table below
Add sterile ddH2O according to the table below
Report your sequencing reactions to Dr. Griffitts or online
Take your sequencing reactions to 690 WIDB and put them in the refrigerator
NOTE: Make sure the liquid is at the bottom of the tube
Recipes
If you load 4 μL of our DNA ladder, the 3 Kb band will represent 12.5 ng/μL. To determine how much DNA to use for sequencing, compare the brightness (thickness) of the 3 Kb band to your DNA band then consult the table below. To sequence 600 to 800 bases, the BYU sequencing center requires 6.67 ng/μL.[1]