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PN 103-140-400 REV05 JUN2024
2b. Custom primer addition for sequencing pre-existing P5/P7 libraries clustered with NO Onso
indexed library control spike-in
Ensure the sequencing reagent pack is thoroughly mixed. Rotate the custom primer addition tubes in the
reagent pack (positions 1-4 as denoted in the image at the start of the section) such that the liquid level in the
tube is visible from the side facing the user.
Using a fresh P1000 pipette tip for each tube, puncture the foil seals of the custom primer tubes in positions
1-4 of the reagent pack. Discard the tips after each use.
Using a P200 pipette, spike in 20 µL of primer solution into the appropriate position of the reagent pack as
indicated in the table at the start of the section, using the following guidelines:
o Spike in 20 µL of the appropriate 100 µM primer stock solution for each custom primer into the
appropriate reagent pack position.
o Visually inspect the side of the custom primer tubes to ensure the primer is added directly to the
buffer in the tube by submerging the pipette tip in the buffer solution during primer addition.
Using a P1000 pipette set to 1000 µL, pipette mix each tube at least 10 times after primer addition. Be sure to
change pipette tips between tubes. Visually inspect the side of the custom primer tubes during mixing to
ensure the pipette tip is submerged in the solution during mixing.
Description of Custom Primer
Custom Insert Read 1 Primer
Custom Index 1 Primer (downstream of the insert)
Custom Insert Read 2 Primer
Custom Index 2 Primer (upstream of the insert)
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