Thermo Fisher Scientific
35
Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Appendix A
Troubleshooting
Table A-1.
Troubleshooting guide
Problem
Possible cause and actions
Low DNA yield
There should be an adequate volume of the Elution Buffer to cover
the KingFisher Magnetic Beads completely during the elution step.
Do not let the KingFisher Magnetic Beads dry, as this may result in
lower elution efficiency.
Heating during the elution step or using a preheated Elution Buffer
enhances the release of the DNA from the KingFisher Magnetic
Beads and the yield of the purified DNA.
Efficient homogenization and lysis of the plant samples increase the
DNA yield.
Prolonged storage of the sample material may reduce the DNA yield.
If the KingFisher instrument mixes too strongly, it may cause partial
elution of DNA already in the Wash Buffer 3.
Use only Thermo Scientific KingFisher plates, tubes and tip combs
with the KingFisher instruments. Use of products from other
manufacturers may cause unsuitable mixing and affect the yield of
purified DNA.
Low purity
Prolonged storage of the sample material may reduce the quality of
the DNA.
Insufficient washing causes impurities in the Elution Buffer.
Carryover of 80% ethanol from the EtOH step of the BindIt protocol
may cause unsatisfactory performance in downstream applications.
Carryover of the KingFisher Magnetic Beads to the Elution Buffer
may affect the A
260
/A
280
ratio. Make sure that the KingFisher Magnetic
Beads do not affect the measurement by centrifuging the samples
or placing them on a magnet for a few minutes to collect the
residual beads at the bottom of the well. Carryover of the KingFisher
Magnetic Beads does not affect most downstream processes.
Continued
