What is the minimum elution volume that can be used with the Monarch® Spin gDNA Extraction Kit columns?

The minimum elution volume is 35 µl. For maximum recovery, 100 µl is recommended. Smaller elution volumes will lead to more concentrated samples but also a lower yield. Typically, when 35 µl is used for elution, yields are approximately 20% lower than when using 100 µl. Using elution volumes lower than 35 µl will result in inconsistent wetting of the silica matrix and significantly reduced recovery.

Optimal elution volume range for elution is 35 – 100 µl. A lysate pool was prepared by using 10 mg RNAlater®-stabilized rat liver samples and following the Monarch tissue protocol. Elution volumes used in triplicates samples were 25, 30, 35, 40, 50, 75, and 100 µl. The average yield obtained with 100 µl elution volume was 19.1 µg, which was considered 100% yield. Using 35 µl to elute, the average yield was 81.6%; 25 µl and 20 µl yielded 74% and 70.6%, respectively.

For volumes ≥35 µl, the recovered volume after elution was ~5 µl lower than the elution volume added and for <30µl the recovered volume was ~10 µl lower. Tests performed with other starting materials showed similar results, with a 20-25% reduction in overall yield when elution volumes were reduced from 100 to 35 µl.

For more information, see “Considerations for Elution and Storage” in the product manual.

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