Home Protocols Standard Cleanup Protocol using the Monarch® Spin PCR & DNA Cleanup Kit and Centrifugation (NEB #T1130)

Standard Cleanup Protocol using the Monarch® Spin PCR & DNA Cleanup Kit and Centrifugation (NEB #T1130)

Buffer Preparation

Prepare buffers as recommended in Buffer Preparation Guidance.


Standard Cleanup Protocol using Centrifugation

  1. Add 5 volumes (e.g., 250 μl) of Monarch Buffer BZ to 1 volume (e.g., 50 μl) of sample. Mix well by pipetting up and down or flicking the tube. Do not vortex. Using a sample volume of 20-100 μl is recommended. For samples less than 20 μl, adjust the volume with TE or nuclease-free water to 20-100 μl. For diluted samples larger than 800 μl, load 800 μl first, proceed with step 2, and repeat as needed.

  2. Insert the Monarch Spin Column S1A into the Monarch Spin Collection Tube and load the sample onto the column. Spin for 1 minute, then discard the flow-through.

  3. Re-insert the column into the collection tube. Wash by adding 200 μl of Monarch Buffer WZ and spin for 1 minute. Discarding flow-through is optional.

  4. Repeat wash (step 3).

  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. For larger size DNA (≥ 15 kb), incubate the column with elution buffer at room temperature for 5 minutes to maximize the yield. Alternatively, heating the elution buffer to 50°C prior to use can be used.