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How can one increase the throughput of the Wizard® Genomic DNA
Purification Kit?
The Wizard® Genomic DNA Purification Kit has been used successfully with
blood sample volumes from 20µl to 10ml (1). Typically for 96 well plates, blood samples
of 20-50µl per well are prepared. For individual tubes, blood samples of 50µl-10ml are
prepared. Processing small volumes of blood (20-50µl) in a 96 well plate will increase
the throughput of the procedure and decrease handling time. Small amounts of blood can
yield DNA sufficient for PCR amplification, but may not produce sufficient amounts for
membrane hybridization (e.g., Southern and dot/slot blots) or digestion with restriction
endonucleases. For example, 50µl of blood will generally yield between 0.2-0.7µg DNA
depending upon the number of leukocytes in the sample.
When using the 96 well protocol for small (e.g., 20-50µl) samples of blood, the DNA is
precipitated across the bottom surface of each well. Larger samples can be processed using
single tubes. For up to 1.0ml of blood, more than 50% of the DNA is rehydrated in the
provided DNA Rehydration Solution after 20 minutes at room temperature compared to
overnight rehydration at 4°C. Both methods produce DNA suitable for PCR.
Reference
- Koller, S., Shiels, G. and Bitner, R. (1999) Using the Wizard® Genomic DNA
Purification Kit with 96 Well Plates. Promega Notes 73, 20.
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