Frequently Asked Questions - PCR Purification

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Both the SOPE resin and the plates should be stored at 4° C.

Expect recoveries of your purified PCR product to be 75%-85% of your unpurified PCR product.

There is no known upper limit to the size of fragment that can be purified. Your recoveries will decrease, however, if you try to purify fragments smaller than 100 base pairs.

No. This product will not remove primer-dimers from your PCR reaction.

Expect recoveries of your purified PCR product to be 75%-99%.

Yes. Use deionized water to bring all reactions <20 µl up to 20 µl.

There is no known upper limit to the size of fragment that can be purified. Your recoveries will decrease, however, if you try to purify fragments smaller than 100 base pairs.

Both the SOPE resin and the cartridges should be stored at 4° C.

The presence of mineral oil in excess amounts will interfere with the performance of QuickStep2. If you use mineral oil, remove as much of it as possible prior to purification.

Yes. Use deionized water to bring all reactions <20µl up to 20µl.

There is no known upper limit to the size of fragment that can be purified. Your recoveries will decrease, however, if you try to purify fragments smaller than 100 base pairs.

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

 

Yes. This product will efficiently remove primer-dimers from your PCR products. We have tested the removal of a 50 base pair fragment.

Yes. Use deionized water to bring all sample volumes ≤50µl up to 100µl.

The minimum volume of PCR product that may be purified on these plates is 50 µl. Samples must be brought up to ≥ 100 µl using deionized water.

a. Yes, but we recommend that you dilute the BigDye Terminator v3.1 to 1 µl or less to insure optimum performance. Be sure to use sequencing reaction diluent, rather than water, to reduce the concentration of BigDye® version 3.1 in your sequencing reaction.

b. If you are using BigDye Terminator v3.1, and experiencing dye blobs, pre-spin the cartridges at 850 x g for 3 minutes, transfer the cartridge to a clean tube, load the sample onto the cartridge and spin at 850 x g for 3 minutes.

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Pre-spin the cartridge at 750 x g for 2 minutes, transfer the cartridge to a clean tube, load the sample onto the cartridge and spin at 750 x g for 2 minutes. If you are using BigDye® Terminator v3.1 and experiencing dye blobs, pre-spin the cartridges at 850 x g for 3 minutes, transfer the cartridge to a clean tube, load the sample onto the cartridge and spin at 850 x g for 3 minutes

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Yes. If you are running 5 µl reactions, adjust the sample volume to at least 10 µl.

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