Promega's Automated MagneSil* ONE, Fixed Yield Plant Genomic
DNA Purification Protocol on the BiomekŪ FX
Promega Corporation – www.promega.com
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version of the following:
| Problem |
Possible Cause |
Solution |
| Low Yield |
- Alcohol present in eluted material
|
- Make sure all Wash Buffer is aspirated before elution.
- Increase drying time on the Heat Transfer Block.
|
|
| Low Yield |
- Insufficient amount of starting material
|
- The optimal amount of starting
material should be determined for each plant species. See
note in Section III.A, Step 1, for guidelines for leaf tissue
samples.
|
|
| Green or Cloudy DNA |
- Too much starting material
|
- Decrease the amount of starting material by 30% and re-isolate
DNA.
- Centrifuge the plate and transfer the supernatant.
|
|
| Green or Cloudy DNA |
- Cold laboratory temperatures may cause reagent precipitation
|
- Centrifuge the plate and trasfer the supernatant.
|
|
| MagneSil* Particles Visible in
Eluate |
- MagneSil* Particle carryover into eluted DNA
|
- Allow more time and mixing on the magnet before removing
eluted DNA.
- Use the minimal amount of MagneSil* Particles necessary
to obtain the desired yield.
- Remove particles by centrifuging the plate and transferring
the supernatant.
|
|
| DNA Too Dilute |
| |
- Decrease elution volume to a minimum of 50µl.
|
|
| DNA Too Dilute |
- Insufficient amount of starting material
|
- The optimal amount of starting material should be determined
for each plant species.
|
|
| Amplification Failure |
- DNA degrades upon storage
|
- Use TE buffer for elution. Store DNA at –20°C
in polypropylene plates or tubes.
|
|
| Amplification Failure |
- Plant may contain phenolics
|
- Add 10mg insoluable PVPP (polyvinylpolypyrrolidone) to
plant material before lysis and grinding step.
|
|
* All trademarks are the property of their respective owners. Where applicable,
the PCR process is covered by patents owned by Roche Molecular Systems,
Inc., and F. Hoffman-LaRoche, Ltd.
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