Griffitts:DNA sequencing

DNA Prep

 * Make a 1:10 dilution of each of your primers
 * 3 μL primer + 27 μL ddH20 will usually suffice
 * NOTE: Each sequence will require two samples&mdash;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 up PCR 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.