User guide

37
Fragment Library Preparation Using the AB Library Builder
™
System: 5500 Series SOLiD
™
Systems User Guide
Chapter5 Troubleshooting
After the automated run
No elution volume Sample volume is lower
than the recommended
volume, leading to wet
filter barrier on the tip
and blockage of nozzles.
In future runs, use the recommended sample volume for the protocol
you are using.
Long-term operation with lower-than-recommended sample volumes
can lead to issues with liquid handling performance.
No amplifiable
library
Insufficient or no
adaptors added to the 5✕
Reaction Buffer tube
Add sufficient adaptor according to the adaptor calculations, and insert
the tube in position 11 of the cartridge (see “Load adaptors in the
cartridges” on page 21).
Enzymes or buffer not at
bottom of wells
Tap the wells down against a hard surface to move enzymes and buffer
to bottom of wells, then inspect the wells.
Observed DNA
peak size is
significantly
different from the
expected DNA peak
size
Incorrect volume in
sheared DNA or
prepared 5✕ Reaction
Buffer tube
Add the correct volumes to the sheared DNA and 5✕ Reaction Buffer
tubes.
Enzymes or buffer not at
bottom of wells
Tap the wells down against a hard surface to move enzymes and buffer
to bottom of wells, then inspect the wells.
Final library is
brownish
Beads in final library 1. Place the tube with the final library in a DynaMag™-2 Magnetic Rack
for at least 1 minute until the solution is clear of brown tint when
viewed at an angle.
2. Without disturbing the pellet, carefully transfer the supernatant,
which contains the final library, to a new 1.5-mL LoBind Tube.
Observation Possible Cause Recommended action