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For PCR products > 300 base pairs, we recommend 20 in Hg. For PCR products 100 – 300 base pairs, we recommend 10 in Hg. To convert to other units, please use our Pressure Conversions Chart.
To convert from Inches
of Mercury (in Hg)
|Millimeters of Mercury (mm Hg)||25.4|
|Pounds per square inch (psi)||0.491153|
We recommend vacuuming the plate for 5 – 10 minutes, or until the wells are empty. The wells of the plate appear shiny when dry. Alternatively, use a pipette to verify if there is any remaining liquid in the wells. Do not over dry the plate. Vacuum times increase when processing > 100 µl volumes.
The wash step is optional. PCR products are suitable for DNA sequencing and microarray spotting after the initial vacuum step. Some sensitive downstream applications may need higher purity DNA and require the optional wash step.
The ExcelaPure 96-Well UF Plate has a standard SBS footprint that is compatible with most vacuum manifolds. We evaluated the following vacuum manifolds during product development:
List of Validated Vacuum Manifolds
|Manufacturer Vacuum Manifold||Manufacturer Catalog Number|
|Millipore MultiScreen Vacuum Manifold||MAVM 096 OR|
|QIAvac Multiwell Unit||9014597|
|Whatman UniVac 3 Vacuum Manifold||7705-0102|
When resuspending your purified PCR product manually, ensure that the product is adequately mixed by vigorously pipetting up and down 20 times. When using a plate shaker, optimize shaker speed and time to ensure sufficient agitation. For liquid handling instruments, optimize aspiration/dispensing of your purified PCR product. Also, check the vacuum setting, we recommend 20 in Hg for PCR products > 300 base pairs. For PCR products 100 – 300 base pairs, we recommend 10 in Hg.