Home Protocols Oligonucleotide Cleanup Protocol for Monarch® Spin PCR & DNA Cleanup Kit (NEB #T1130)

Oligonucleotide Cleanup Protocol for Monarch® Spin PCR & DNA Cleanup Kit (NEB #T1130)

Buffer Preparation

Prepare buffers as recommended in Buffer Preparation Guidance for Monarch Spin PCR and DNA Cleanup Kit 5 ul.

 

Oligonucleotide Cleanup Protocol

  1. Add 100 μl Monarch Buffer BZ (2 volumes) (ensure that isopropanol has been added, as indicated on the bottle label) to 50 μl (1 volume) sample. A sample volume of 50 μl is recommended. For smaller samples, adjust the volume with nuclease-free water. If using a sample > 90 μl, reloading the column during step 3 will be required.

  2. Add 300 μl (2 volumes) isopropanol. Mix well by pipetting up and down or flicking the tube. Do not vortex.

  3. Insert the column into the collection tube and load the sample onto the column. Spin for 1 minute, then discard the flow-through.

  4. Re-insert the column into the collection tube. Wash by adding 500 μl Monarch Buffer WZ and spin for 1 minute. Discard flow-through. Additionally, a second wash step with Monarch Wash buffer WZ or 80% ethanol can be carried out to ensure the removal of enzymes that may interfere with downstream applications.

  5. Transfer the column to a clean 1.5 ml microfuge tube. Use care to ensure that the tip of the column does not touch the flow-through. If in doubt, re-spin for 1 minute.

  6. Add 5-20 μl of Monarch Buffer EY to the center of the matrix to elute DNA. Wait for 1 minute, and spin for 1 minute. Nuclease-free water can also be used to elute the DNA. Yield may slightly increase if a larger volume of Monarch Buffer EY is used, but the DNA will be less concentrated.