1. If using custom sequencing primers, load them in the correct wells on the reagent cartridge the following way:

  1. a. Find the appropriate wells for your custom sequencing primers, which are marked by the green sticker, and are used in the following order:

    1. ACustom read 1 primer
    2. BCustom read 2 primer
    3. CCustom Index 1 primer
    4. DCustom Index 2 primer
  2. b. Pierce the foil of the wells to be used with a DNAse-free 1 mL pipette tip.
  3. c. Load 2 mL of each 1 µM custom sequencing primer in the appropriate wells.

Note

If you need to dilute the custom sequencing primers, make sure to use the 5X Primer Annealing Buffer (5X PAB) to arrive at a final buffer concentration of 1X PAB. For example, for diluting a 100 µM primer stock to 1 µM final concentration, mix 1580 µL molecular-grade water, 400 µL 5X PAB, and 20 µL primer stock, and load in the custom sequencing primer well.

2. Make sure to specify in the sample sheet that you are using custom sequencing primers.