If a variable speed centrifuge is unavailable, a fixed speed centrifuge can be used. However, reduced yields may be observed.

Varying the centrifugation speed from the recommended setting could result in a decrease in oligonucleotide yields.

Yes, you can. All the solutions included in the sample preparation step of this kit are in a linear relationship to the volume of oligonucleotide sample processed. Make sure that you do not deviate from the ratio specified in the product manual. The solutions are optimized per 50 µL of oligonucleotide sample.

Adding less volume may reduce your oligonucleotide yields. Adding more may not affect the oligonucleotide yields EXCEPT if more Elution Solution A was added.
Eluting oligonucleotide in more Elution Solution will result in diluting your yield.

If you forget to perform a dry spin after the third wash, your oligonucleotide elution may become contaminated with Wash Solution A. This has the potential to dilute the oligonucleotide yield in your elution and could also interfere with downstream applications.

If the recovery of the oligonucleotide is poor, ensure that isopropanol was added during the sample preparation step and ethanol was added to Wash Solution A. Additionally, it is recommended to use the Elution Buffer provided with the kit for optimal recovery.

The underperformance of your oligonucleotide in downstream applications may be attributed to the following factors:

• Insufficient washing of the column.

  Ensure that the column is thoroughly washed three times with Wash Solution A during the binding step to eliminate any traces of salt, as residual salt can hinder downstream applications.

• Incomplete removal of ethanol.

  Make certain that the dry spin under the Column Wash procedure is executed to remove ethanol traces before elution, as ethanol can interfere with various downstream processes.

• Use of a different Elution Solution.

  If a different Elution Solution was used instead of the provided kit solution, assess it for components that might disrupt the application, such as high salts (including EDTA), detergents, and other denaturants. Verify the compatibility of your chosen elution buffer with the intended downstream use to address any potential interference issues.