- Materials
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- Monarch® DNA Capture Beads
- Monarch® Bead Retainer
- Monarch® Collection Tubes II
- Consumables
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- 2 ml Eppendorf DNA LoBind tubes
- Qubit dsDNA BR Assay Kit (Invitrogen, Q32850)
- Equipment
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- Vortex mixer
- Centrifuge
- Qubit fluorometer (or equivalent)
- P200 pipette and tips
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Quantification of uHMW gDNA
The method to quantify uHMW gDNA was developed by Paul A ‘Giron’ Koetsier & Eric J Cantor, 2021, which recommends the use of a regular P200 pipette and tip.
This optional uHMW gDNA quantification step has also been included in the protocol for user QC. However, this step can be omitted and 750 µl of DNA in Extraction EB (EEB) can be taken straight into the tagmentation step of the protocol.
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Use a regular P200 pipette tip to aspirate 10 µl of gDNA.
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Dispense the aspirated gDNA into a fresh 2 ml Eppendorf DNA LoBind tube.
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Add a Monarch DNA Capture Bead to the 10 µl of gDNA and vortex aggressively for 1 minute to shear the gDNA.
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Transfer the gDNA and beads into a clean Monarch Bead Retainer inserted in a Monarch Collection Tube II. Spin the tube at 1000 x g for 1 minute to separate gDNA from the beads. Dispose of beads and bead retainer.
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Transfer the gDNA into a clean 1.5 ml Eppendorf DNA LoBind tube.
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Quantify the sample using a Qubit fluorometer. The expected yield is 30-40 µg of DNA.