If the final sample Tapestri library exhibits low-size off-target products (e.g., 200 – 240 bp) that make up more than 5 % (and less than 50 %) of all products based on molarity (number of molecules), then follow the steps outlined below.
After eluting 10 µL of Library PCR product after Ampure XP cleanup and using 1 µL for subsequent BioA assay, follow these steps:
-
Add 16 µL of nuclease-free water to 9 µL sample for a total volume of 25 µL.
-
Mix and quick-spin to collect the contents.
-
Add 18 µL (0.72x) of Ampure XP reagent, at room temperature and well-mixed, to the above sample.
-
Vortex for 5 seconds and quick-spin to collect the contents.
-
Incubate the tube at room temperature for 5 minutes.
-
Place on the magnet and wait 5 minutes for the beads to separate from the solution.
-
Without removing the tube from the magnet, remove the clear liquid from the tube and discard.
-
Add 100 μL of the freshly prepared 80 % ethanol, wait 30 seconds, and remove 100 μL of ethanol without disturbing the Ampure beads.
-
Repeat Step 8 once for a total of two wash cycles.
-
Remove all residual ethanol from the tube. Take the tube off the magnet and do a quick spin. Place the tube back on the magnet with the caps open and remove any residual ethanol.
-
Dry the Ampure bead pellets in the tubes on the magnet by incubating at room temperature for 2 – 5 minutes. Avoid overdrying the beads.
-
Remove the tube from the magnet. Add 9 μL of nuclease-free water into the tube. Vortex and quick-spin to collect the contents.
-
Incubate the tubes at room temperature for 2 minutes.
-
Place the tube onto the magnet and wait for at least 2 minutes or until the solutions are clear.
-
Transfer 8 μL of purified PCR product from the tube to a new 0.2 mL PCR.
If the final sample Tapestri library exhibits low-size off-targets (e.g., 200 – 240 bp) that make up more than 50 % of all products based on molarity (number of molecules), contact Mission Bio Support at support@missionbio.com.
